Cytotherapy (2001) Vol. 3, No. 1, 55–60

55

Fourth International Symposium on HematopoieticStem Cell Transplantation

The Institute of Medical ScienceThe University of Tokyo (IMSUT), Tokyo, Japan

July 5–6, 2000

© 2001 ISHAGE

The Fourth International Symposium on HematopoieticStem Cell Transplantation was held on July 5–6, 2000 at theInstitute of Medical Science, Tokyo University. TheSymposium was organized by the Science Council of Japan,under the direction of Drs Keisuke Toyama and S.Asano. This Symposium has been widely attended by Asianstem-cell scientists and transplant physicians.

The abstracts of the Symposium include clinical reportsfrom the Placental Blood Program at the New York BloodCenter, details of the activity of the International NET-CORD Organization, and the first clinical report of cord-blood transplantation in a nuclear accident patient. Morethan 200 experts attended this meeting and intensive dis-cussion followed.

Each Symposium theme has reflected the increasingreliance on cord blood as a source of hematopoietic stemcells in Asia:n (1996) Present status of cord-blood stem-cell transplan-

tationn (1998) Responding to diversity and information

exchange in cord-blood bankingn (1999) Network of cord-blood banks worldwiden (2000) Future of cord-blood stem-cell transplantation.Symposium speakers included: Drs P Rubinstein, C Stevens(New York, USA), E Gluckman (Paris, France), G Sirchia(Milan, Italy), P Wernet (Dusseldorf, Germany), JA Madrigal (London, UK), J Fraser (Los Angeles, USA), L Harvath (Bethesda, USA), P Coelho (Rancho Cordova,USA),DLu(Beijing,China),ZHan(Tianjin,China), KH Lin

(Taipei, Taiwan), H Han (Seoul, Korea), S Issaragrisil(Bangkok, Thailand), Tran van Binh (Ho Chi Minh City,Vietnam) and leading stem-cell scientists from Japan.

The increasing importance of cord blood in Asia isdemonstrated by the fact that the Japan Cord Blood BankNetwork has supplied 283 grafts to patients and has bankedmore than 4000 units. Approximately 7000 units are nowbanked in China, where there is an urgent need for cord-blood banks because a policy of having fewer childrenmakes it difficult to match BM donors to patients.

At the Third Symposium in 1999, a network of cord-blood banks in Bangkok (Thailand), Ho Chi Minh City(Vietnam), Seoul (Korea), Taipei (Taiwan), Beijing (China),Tianjin (China) and Tokyo (Japan) was formed and namedAsiaCord, with its secretarial office at the Tokyo CordBlood Bank. The scientific committee is now developing astandard (minimum requirements) for cord-blood banking,to secure the quality and safety of the units stored in thenetwork. The AsiaCord web site, which is intended to assistthe search for units, is under preparation.

Many of the cord-blood banks around the world havedifficulties with finance and AsiaCord is no exception.Nevertheless, the goal of AsiaCord is to supply high, con-sistent-quality cord-blood units to patients in Asia.

The Fifth Symposium will be held on June 22 and 23,2001 at the Institute of Medical Science, The University ofTokyo, Japan. The theme will be ‘Placenta/umbilical cordblood and regeneration biology’. We invite all of you whowish to attend the Symposium to contact the address below.

Fourth International Symposium on Hematopoietic Stem Cell Transplantation

Hematopoietic stem cell transplantationMeeting Report

TA Takahashi

Correspondence to: Tsuneo A. Takahashi, Division of Cell Processing, The Institute of Medical Science, The University of Tokyo,Shirokanedai 4-6-1, Minato-ku, Tokyo, Japan.

56© 2001 ISHAGE

Abstracts 57

JAPAN CORD BLOOD BANK NETWORKJ Kato, Japanese Red Cross Nagoya First Hospital, Nagoya, JapanSince the success of unrelated cord blood transplantation (CBSCT) in 1993 in theUSA, cord blood (CB) has been increasingly used as a source of hematopoieticstem cells for transplantation. In Japan, the first CB bank was established in 1995in Kanagawa and nine banks were established by 1999. In August 1999, the JapanCord Blood Bank Network was organized, with local CB banks, and was funded bythe Ministry of Welfare. It is expected that 20 000 CB units will be collectedwithin 5 years to supply HLA 6/6 or 5/6 matched CB units to over 90% ofpatients.

Since 1997, 202 cases of unrelated CBSCT were undergone for patients withmalignant diseases (n = 168) and non-malignant diseases (n = 34). Patients were0–50 years old (median 6 years) and 5.3–69 kg body weight (median 19 kg).Engraftment was evaluated in 170 transplants and was 83.7% for malignantdiseases, and 62.5% for non-malignant diseases. Grade II–IV acute GvHD wereobserved in 38% of all patients and HLA disparity did not affect the developmentof acute GvHD. DFS of patients with HLA 0–1 and 2–3 Ag-mismatched graftswere 40.2% and 45.4% in malignant diseases, and 28.2% and 33.3% in non-malignant diseases, respectively. DFS of patients with malignant diseases atstandard risk was 72.0% and that of patients at high risk was 22.5%. For malignantdiseases, children < 5 years of age and with graft of > 4 x 107/kg infused cells hadsignificantly better DFS. Causes of death were mostly infection, relapse andtreatment-related complications.

The results of unrelated CBSCT in Japan were comparable to those of previousreports from the USA and Europe, in terms of engraftment, GvHD, DFS andcauses of death. The unrelated CBSCT is a feasible procedure for certain patients,especially children with malignant diseases. More extensive studies with a largernumber of patients will elucidate the long-term outcome of unrelated CBSCT.

IMMUNOLOGICAL PROPERTIES OF CORD BLOOD CELLSS Cohen, A Madrigal, The Anthony Nolan Research Institute, Royal FreeHospital and University College Medical School, Pond Street, Hampstead,London, NW3 2QG, UKIt is generally agreed that cord blood (CB) transplantation represents anencouraging alternative BMT. A variety of observations have been made to explainthis, the two most controversial being 1) there is less GvHD with CB comparedwith BMT and 2) more HLA mismatches can be used with CB transplantation. Ifthese two factors are true, then CB may generate a lower ‘immunological response’compared with transplantation with adult cells. Why this may be is unknown,however, much work has been done recently to compare the immunologicalfunction of CB and adult monocytes. This work may begin to explain some of thedifferences we see with different transplants. The most common explanation forreduced CB cell function is that it is naive, since, compared with adultmononuclear cell populations, there are (a) higher levels of CD4+CD45RA+ cells,(b) lower Ag and mitogen specific T-cell proliferation, (c) lower cytokineproduction on stimulation and (d) a more polyclonal T-cell receptor repertoirecompared with adult blood. However, lymphocyte naivety may not be the onlyanswer since it is possible these naive phenomena, noted in vitro, are bypassed invivo. We have evidence that there are differences in the soluble factors secretedinto the serum that altered the function of cord and adult lymphocytes, and thatthere may be active suppression of CB natural killer (NK) cells.Role of soluble factors We have analysed the effect of heat-inactivated sera onthe function of T-cell proliferation by looking at the mitogen- and cytokine-specific proliferative responses. Using adult PBMC and both human and mouse T-cell lines, we have shown that adult sera will enhance the proliferative responses,whereas CB sera has no effect. These results suggest that there is a factor withinmature serum that is absent in fetal serum. This factor is, in part, responsible forthe proliferation of T-cells and its absence could explain the reduced proliferativeresponses of CB compared with adult T-cells.Suppression of CB NK cells Previous studies have reported reduced NK cellactivity in CB compared with adult blood mononuclear cell populations. Using anon-radioactive killing assay, we have verified these findings, suggesting that eitherthe fetal NK cell function is suppressed, or that these cells are functionallyimmature. We have shown that CB NK cells are functional, since activating themwith cytokines known to activate adult NK cells (IL2, IL12 and IL15) increasedactivation. However, resting the cells, which enhanced adult NK cell activity (p <0.01) had no effect on fetal NK cells (p = 0.2). These results suggested that fetalNK cells have the capacity to kill, but this is suppressed in vitro. This hypothesiswas strengthened by our observation that 8/9 CB mononuclear cell populationshad their NK activity restored by freeze/thawing, whereas 4/5 adult PBMC had areduced killing ability on freeze/thawing, or they are suppressed in situ. Thus,either NK cells are activated to kill by the freeze/thawing. ??? To determine whichwas the case, we performed extensive phenotypic analysis of the CB populations

pre- and post-freezing, and found that the percentage of CD3-CD56+ populationwithin CB increased significantly (p < 0.0005 by paired t test) with freezing,whereas freeze/thawing had no effect on this population within normal adultPBMC population. Our data suggests that within CB there is a population of cells,as yet undefined, that may be inhibiting NK function.

In conclusion, we have shown that there are factors within the CB that inhibitlymphocyte function, either by a lack of a soluble factor, or by active suppressionof NK cell function. These results provide suggestions as to why CBtransplantation may give a lower GvHD than BMT and help us begin tounderstand the mechanisms involved in maternal tolerance to the fetus inpregnancy.

EFFECT OF TRANSIENT WARMING EVENTS ON CELL VIABILITYOF PLACENTAL CORD BLOODP Coelho1, L Dobrila2, and P Rubinstein2, 1ThermoGenesis Corp. and 2NewYork Blood CenterBackground The viability of cryopreserved hematopoietic stem and progenitorcells from placental/umbilical cord blood (PCB) units may be reduced as a resultof the transient warming events (TWEs) that affect them after initial freezing.These TWEs occur during routine post-freeze processing, storage and shipping ofPCB grafts. Units are exposed to the ambient air as they are transferred from thecontrolled-rate freezer to quarantine freezer and from quarantine to storagefreezer; during storage, as different units stored in the same rack are removed;while being moved from storage to cryo-shipper; and at least once more when theunit is placed in the storage dewar?? at the transplant center.Methods The volume of three PCB units was reduced to 20 mL by removing mostof the red cells and plasma, and then cryoprotected (10% DMSO), separated intoaliquots of 1 mL, frozen at 2°C/min to -80°C and placed into a liquid nitrogen(LN2) tank. Quadruplicate vials containing these aliquots were then transferred toalcohol baths maintaining -25°C, -40°C or -80°C. They were kept at thesetemperatures for 4 min and then returned to LN2. After 5 min LN2, two aliquotsthat had been exposed to each of these temperatures were returned to the samealcohol baths and underwent five identical cycles, from LN2 to -25°C, -40°C and-80°C, respectively, and put back into LN2. A control group of four aliquots wasmaintained at -196°C. All aliquots of PCB were then thawed, the DMSO waswashed from the cells, and examined by acridine-orange and ethidium bromidestaining for recoveries of total and viable leukocytes. The recoveries of viableprogenitor cells were ascertained by (FACS) CD34 cell counts.Results Total and viable leukocyte recoveries were calculated as the differencebetween the recovery from aliquots kept in LN2 throughout and those exposed toTWEs either once or five times. Total cell recoveries were 97–102% of controls inall cases. The viabilities of the cells exposed to TWE are summarized in Tables 1and 2.Conclusionsn TWEs can cause measurable decreases of cell viability, including in CD34+

mononuclear cells.n The damage encountered in these experiments appears to depend on the

magnitude of warming and to be cumulative when the TWE is repeated fivetimes.

n The effects are being tested in parallel studies in which viability is ascertainedby progenitor cell cultures, to determine the specific susceptibility of theCFU-C populations.

n It is perhaps possible that a single exposure to -80°C may not producesignificant enough damage to warrant the expense of shipping frozen PCBgrafts at LN2 temperatures. This possibility is also being tested by repeatedexperiments in which the effect of variable times of exposure to -80°C and -70°C is being studied alone and in combination with other TWEs.

CORD BLOOD TRANSPLANTATION FOR ADULT PATIENTS,INCLUDING NUCLEAR ACCIDENT PATIENTT Iseki, Division of Molecular Therapy, Advanced Cancer Research Center,The Institute of Medical Science, The University of Tokyo (IMSUT)Although the number of cord blood transplants (CBT) has been rapidly increasing,most of the reported patients are children. Several reports from America andEurope described relatively high mortality rates among CBT patients as a resultof infection. It has been suggested that low cell content and delay in engraftmentmay restrict the use of cord blood in adults. The important clinical issues for adultpatients, such as patient eligibility, optimal cell doses and the proper regimen ofconditioning and GvHD prophylaxis, have not yet been defined. I reported on ourexperience with CBT in adult patients and the first case of CBT in the treatmentof the victim of a nuclear accident.

Between August 1998 and May 2000, 15 adult patients with hematologicalmalignancy received CBT from HLA mismatched unrelated donor at IMSUT.

58 Fourth International Symposium on Hematopoietic Stem Cell Transplantation

Median age of patients was 41 years (16–51), weight was 53 kg (40–68) and thenumber of nucleated cells infused was 2.25 3 107 kg (1.2–4.1). All patients receivedthe standard conditioning regimen of IMSUT according to disease status. Allpatients received CsA and 11 received short-term MTX for GvHD prophylaxis.No patients received ATG. All cases received G-CSF administered after CBT.

Three patients died shortly after CBT (27–39 days), because of RRT (two) andinfection (one). The patient with ALL received full TBI and cytoxan anddeveloped autologous recovery. Among 11 evaluable patients, median time toneutrophil and platelet engraftment was 24.5 days (9–41) and 50 days (35–164, n =9) respectively, which appears to be faster engraftment than previously reported,despite concomitant use of MTX. Three patients developed Grade II and onedeveloped Grade III acute GvHD. Three patients died of relapse 107–307 daysafter CBT. Overall survival rate at 22 months was 40 ± 19% for all patients and 53± 23% for 11 patients in the first transplantation group. Survival rate of poor riskgroup (n = 11) was low (33 ± 17%), but all four patients with standard risk are alive70–270 days after CBT. Although the number of patients and periods ofobservation are insufficient, these results are comparable to those from standardBMT.

A 39 year old male was systemically irradiated during a nuclear processingaccident in October 1999. Radiation dose was estimated at around 8 gy equivalent.A severe and prompt decrease of lymphocyte count was observed, and extremelyhypocellular BM, without hematopoietic progenitor cells, was ascertained byrepeated examination from different sites. Although the possibility of unevenradiation exposure and subsequent autologous hematopoietic recovery wasconsidered possible at this point, it was judged that a hematopoietic stem celltransplant was necessary. An HLA identical sibling donor was not available. Therisk of GvHD, which could cause organ damage worse than the radiation damage,was considered less likely to occur with CBT. Furthermore, mixed chimeric phaseand late graft rejection can be obtained relatively easily after CBT compared withother stem cell sources. We therefore selected CBT as the stem cell source.However, because hematological recovery after CBT is regarded as generally slow,we decided to cope with this risk by strict prevention of infections andintroduction of intensive supportive hematopoietic factors, with TPO in additionto G-CSF. ATG, methylprednisolone and CsA were administered for conditioningand GvHD prophylaxis. To avoid additional damage to organs and residualautologous hematopoietic stem cell, if any, we used no cytotoxic agents, such ascytoxan or MTX.

Recovery of neutrophils was observed on Day 16 after CBT, followed byreticulocyte and platelet recovery. No significant infection was observed.Engraftment was ascertained by chromosome analysis on Day 9 after CBT. SerialFISH analysis revealed that mixed chimerism was established shortly after CBT,following which the ratio of graft gradually decreased and was absolutely rejected3 months after CBT, when autologous hematopoietic recovery was achieved. Thepatient died of multiple organ failure caused by radiation injury 7 months after theaccident. No clinical or pathological evidence of transplantation-related toxicityor GvHD were noted through his whole clinical course. CBT was considered tobe successful in the sense of rescue during BM aplasia without any adverse effect.

VOLUNTEER ACTIVITIES TO SUPPORT THE NATIONAL CORDBLOOD BANK IN JAPANM Arita, Representative, Group of Volunteers to Support the Cord BloodBank in JapanThe main features of the activities to support the National Cord Blood Bank inJapan lay in the fact that citizens moved the politicians and government, by givingsupport to the promotion of medicine, as well as to research on UC blood cells.This support activity was launched in 1992 with the aim of complementing theactivities of the National Marrow Donor Registry (founded in 1991). Until about1994, we publicized the need for UC blood and raised the number of advocates forthis issue, by providing researchers, doctors and medical journals with informationobtained through investigation on cord-blood banks overseas, and withinformation on cord-blood stem-cell transplantation. Meanwhile, in the practicalsphere of medical service/research, local cord-blood bank facilities wereestablished one after another after 1995, centering around the activity of ourvolunteer group. With the technical progress in collection, separation andcryopreservation of UC blood cells, the clinical outcome of cord-blood stem-celltransplantation in Japan rapidly proved to be a success. Following visits to the CordBlood Bank in New York Blood Center in 1996, and Duke University the followingyear, we learned that cord-blood stem-cell transplantation could be a potentialremedy for patients with different types of illnesses. In order that these potentialremedies could become realities, we decided that our activities to support thecord-blood bank would be developed further, into a social activity of wider span.Japan has a system of health insurance for the whole nation. In 1997, we started asignature-collecting campaign, appealing for the application of medical insurance

to cord-blood stem-cell transplantation and the founding of a cord-blood bank. Wesubmitted a total of 1.5 million signatures directly to the Minister of Health andWelfare, and the Speaker of the House of Representatives and the President of theHouse of Councillors. By March 1998, a total of 2.2 million signatures had beendirectly submitted to the Minister. The signature-collecting campaign waspolitically supported by more than 440 municipal assemblies in their submissionof opinion to the government and question in the Diet. In 1998, medical insurancewas applied to cord-blood stem-cell transplantation and in 1997 a national cord-blood bank was founded, incorporating eight local networks of blood banks andrealizing services for patients based upon a principle of competition.

Specific volunteer support activities for the cord-blood bank include:transportation of cord blood from obstetric institutions, appealing to pregnantwomen to provide cord blood, as well as various donation activities to the cord-blood banks. The three pillars of future volunteer activities to support the nationalcord-blood bank are as follows.n Assistance in child-raising, in terms of providing an environment in which

women are capable of childbirth and child-raising. In more concrete terms,public educational activity to protect youth from environmental pollution anddrug pollution; this will eventually result in pregnancy and childbirth inhealthy condition.

n Support services, in terms of raising the quality of cord blood provided topatients. For example, applying health insurance to umbilical core blood as amedical material, in order to stabilize the operation of cord-blood banks.

n Medical service support, by setting up a large-scale transplantation center thatcan provide prompt transplantation treatment according to patients’ needs,regarded as one of the merits of cord-blood stem-cell transplantation.

Under the present allocation of medical transplantation facilities in Japan, whereoperations are incline to be inadequate, we must be fully aware of the fact that aconsiderable number of patients who need marrow transplantation/cord-bloodstem-cell transplantation are kept waiting too long and denied immediatetransplantation.

OUR EXPERIENCE WITH STEM-CELL TRANSPLANTATION FORMETABOLIC DISEASEH. Mugishima, Nihon University School of Medicine Department ofPediatrics, Tokyo, JapanHematopoietic stem-cell transplantation (HPSCT) as a form of therapy forlysosomal storage diseases primarily serves to replace abnormal cells with theirnormal equivalent, which will then act as a source of the normal enzyme. Thisstrategy relies on adequate secretion of the normal hydrolase from hematopoietic-derived cells and the receptor-mediated uptake of the enzyme by diseased cells.Allogeneic HPSCT from HLA-identical donors has been used with increasingfrequency to treat patients with lysosomal storage diseases. The effects of HPSCTlargely depend on the type and stage of the disease. Generally, visceral symptomscan be improved by transplantation, but skeletal lesions are relatively unaffectedby transplantation. The effect on neurological symptoms varies. HPSCT (fiveusing BM, two using cord blood) was performed in six patients with Huntersyndrome (four), Morquio syndrome (one) and Adrenoleukodystrophy (one)between September 1996 and June 2000 in our institution.The donor was an HLA-identical sibling in two cases, an HLA-identical volunteerunrelated donor in three cases and an HLA-nonidentical volunteer unrelateddonor in two cases. We reported the clinical outcome of these patients.The long-term therapeutic effects of HPSCT can be subject to multiple factors,including a level of biochemical improvement, a reversibility of affected tissues, ora degree of advanced disease prior to HPSCT. Thus, an accumulation of clinicaland biochemical follow-up study is needed to understand the therapeutic efficacyand consequence of HPSCT for lysosomal storage diseases.

PLACENTAL BLOOD FOR UNRELATED BM REPLACEMENT:A SINGLE BANK’S UPDATEP Rubinstein, CE Stevens, New York Blood Center, NY USABackground and objectives Placental/UC blood (PCB) has been used forallogeneic BM replacement for both related and unrelated transplantation since1988. The Placental Blood Program of the New York Blood Center has providedover 1000 unrelated grafts since 1993 and previous analyses of the outcomes ofthese transplants identified variables associated with clinical outcomes. We updatethe information on the clinical outcomes of these transplants through the end of1999, with emphasis on the influence of HLA matching.Material and methods All 932 patients transplanted through 1999 with PCB fromour Program are included in this report. Two-thirds were diagnosed with leukemiaor lymphoma, 25% with inherited conditions and 7% with acquired diseases. Atleast 1 year post-transplant follow-up data was available on > 90% of patientstransplanted 1 year or longer before the set was closed.

Abstracts 59

Results Engraftment was ultimately achieved by 92% of recipients(Kaplan–Meier estimate) and the variable identified earlier as affecting theprobability and speed of engraftment were confirmed. Multivariate analysis alsoconfirmed the independent influence of cell dose, HLA matching, diseasediagnosis and transplant center location (US versus foreign). Patient age and HLAmatch grade independently affected the frequency and severity of acute GvHD.Leukemia relapse was associated with the stage of disease at transplantation andthe prior existence of acute GvHD. The probability of transplant-related eventswas independently associated with disease diagnosis, cell dose, number of HLAmismatches and transplant center, while the cell dose failed to associatesignificantly with the relative risk of reaching this endpoint in the subset ofpatients who achieved engraftment. Overall, event-free survival rates at 1 yearpost-transplant were 49% and 30%, respectively for genetic disease andhematopoietic malignancies and 35% for patients with acquired diseases.Conclusions These results confirm and extend earlier data, particularlyestablishing the significant association of transplant success with histcompatibilitymatching grades and indicating the urgency of improving the transplant matchlevels, by active cooperation among placental blood banks and transplant centersto improve patient survival.

NUCLEATED RBC IN PLACENTAL/UC BLOOD FOR UNRELATEDBM RECONSTITUTION: AN EPIDEMIOLOGICAL PERSPECTIVECE Stevens, P Rubinstein, New York Blood Center, New York, USABackground By June 30, 2000, the New York Blood Center’s Placental BloodProgram had collected 10 849 and provided 1044 fully tested placental/UC blood(PCB) grafts to 1022 patients.Methods PCB units were selected for specific patients based on HLA match andnuclear blood-cell content. Total nucleated blood cells were enumerated by anautomated hematology analyzer and hematopoietic progenitors were assayed bycolony culture. Nucleated RBC number was determined retrospectively by visualinspection of cord blood smears under light microscopy. WBC number wascalculated by adjusting the total nucleated cell (TNC) count for the population ofnucleated RBCs. Mothers who donated PCB units were interviewed and theirmedical records reviewed for their ethnic background, risk factors for infectiousand genetic diseases, and for complications of pregnancy, labor and delivery.Patients’ post-transplant outcomes were based on reports by their respectiveTransplant Centers.Results Nucleated RBC number in PCB was related to total nucleated andprogenitor cell numbers and less well to WBCs. The timing of myeloidengraftment correlated with nucleated RBC dose, as well as with TNC, WBC andprogenitor dose per kg of body weight at the time of transplantation. In amultivariate analysis, both WBC and nucleated RBC dose were predictive ofengraftment rate. When the analysis included progenitor cell dose, only progenitordose predicted engraftment rate. The number of nucleated RBCs inplacental/UCB correlated with factors known to be associated with fetal hypoxia,blood group incompatibility, maternal fever during labor, maternal diabetesmellitus and varied by the infant’s ethnicity.Conclusion A high population of nucleated RBCs in pre-freeze PCB units did notreduce engraftment potential, since nucleated RBC number correlated directlywith hematopoietic progenitor cell number and with speed of engraftment. Factorsthat promote the production and/or release of RBC precursors into the fetalcirculation also may promote production and /or release of hematopoietic stemcells.

THE INTERNATIONAL NETCORD ORGANIZATION AND ITS ROLEIN CORD BLOOD BANKING AND ALLOCATIONP Wernet, Heinrich Hine University, Dusseldorf, Germany (for the Board ofDirectors of NETCORD)In 1998 nine large cord-blood banks founded the International NETCORDOrganization in Italy, in order to provide a direct service of high product qualityto the stem-cell transplantation hospitals, together with some guidance on the bestallocation chances via a joint cord-blood banking inventory through a virtualoffice employing the Internet.

In 2000 NETCORD published, together with FACHT (Foundation for theAccreditation of Hematopoietic Cell Therapy), the First Edition of InternationalStandards for Cord Blood Collection, Processing, Testing, Banking, Selection and Release.These Standards have been adopted by ISHAGE, ASBT (American Society forBlood and Marrow Transplantation), EURO-ISHAGE (European InternationalSociety for Hematology and Graft Engineering), EMBT (European Group forBlood and Marrow Transplantation) and by JACIE (Joint AccreditationCommittee of ISHAGE-Europe and EBMT). They were submitted jointly to theFDA (Food and Drug Administration of the USA) this spring.

The plan is that this will be followed by a joint FACH-NETCORD-

Accreditation Process, based on the joint NETCORD-FACHT AccreditationManual for cord-blood banks worldwide. Meanwhile, NETCORD has 14 memberbanks, with a total of inventory of 46 397 available cord blood (CB) units ready fortransplantation. So far 1641 CB transplantations have been performed worldwidefrom this inventory; 1246 to children and 376 to adults.

Future activities will focus on the completion of the Internet service and on thejoining of many other highly-qualified CB banks, in order to reach the stem-celltransplant physician as fast as possible with the most appropriate CB unit for everypatient worldwide.

CORD BLOOD BANKING, TRANSPLANTATION AND THERATIONALE OF INTRA-AORTIC INFUSIOND-P Lu, K Liu, D Li, H Ren, X Huang, Y Zhang, K Sun, Peking UniversityInstitute of Hematology, Beijing, 100044, PR ChinaCord-blood banking has been moving on smoothly, although slowly, in China.There are cord-blood stem/progenitor cell banks in Beijing, Shandong,Guangdong, Tianjin and Sichuan, storing about 4000, 1000, 1000, 600 and 500cord-blood units respectively. The total accumulated number of cord blood unitsis about 7000 units. They are all HLA-typed by molecular (SSP/SSOP) andserological methods.

Cord-blood transplantation has been successful so far at our Institute usingrelated and unrelated frozen cord blood for patients weighed up to 75 and 95 kgbody wt. The latter received two units of cord blood at the same time. The numberof nuclear cells infused range from 1.73 to 12.0 3 10/kg. The follow-up time isfrom 2 to 16 months. All of the patients are alive.

The route of intra-aortic infusion has been adopted for every unrelated cordblood and BMT. There was engraftment in all patients (n = 14). A mouse animalmodel was used in this present study showing confiscation of hemopoietic cells inpulmonary circulation/lung tissue. Homing of hemopoietic cells in the BM wasstatistically higher in the aortic infusion group than in the i.v. group (at 30 h witha localization index of 14.52 versus 10.49).

CORD BLOOD BANKING AND CLINICAL APPLICATION INTAIWANK-H Lin1, B-W Chen2, J-W Chen3, K-S Lin3, 1National Taiwan University,Taipei, 2Sun Yat-Sen Cancer Center, Taipei; and 3Chinese Blood ServicesFoundation, TaiwanFrom 1993 to 2000, the number of entries in the marrow donor registry hasincreased to about 200 000 donors, among which the probability of finding acomplete match is estimated to be 60%. Three cord-blood banking programs(CBBP) started in 1998, established by Sun Yat-Sen Cancer Center (SYSCC),Chinese Blood Services Foundation (CBSF), and Tsu-Chi Foundation (TCF).

Between December 1998 and May 2000, the CBBP at the SYSCC banked > 500cord-blood units. About the same time, > 700 cord-blood units were collected,processed, HLA-typed, tested for transmissible diseases and cryopreserved usingthe method of the New York Blood Center at CBSF. The CBBP at TCF wasestablished and beginning to collect cord blood.

Among 4% of cord-blood units collected at SYSCC, 24 are not eligible for use.Fresh cord-blood units contain 10.2 x 108 nucleated cells and 8.4 x 108 afterprocessing. HLA is typed on all qualified units with One Lambda ABDR 96-wellSSP DNA tray. The cryo-units contain 2.3 3 106 CD34+ cells, 7.4 3 108 NC, 3.03 108 MNC and 8 3 104 CFU-GM. Among 600 cord-blood units collected atCBSF, the average number of cord-blood units is 1.0 3 109 NC, 3.5 3 106 CD34+

cells, and 4.2 3 105 CFU-GM.The first unrelated cord-blood transplantation (CBT) in Taiwan was performed

for a male infant with Hunter’s disease at National Taiwan University Hospital(NTUH) in June 1999. The donor was a female newborn cord-blood unit collectedat NTUH and processed at CBSF. No engraftment was documented after CBT inthis infant, thus a second transplant was done with a cord-blood unit from SYSCCin January 2000 at NTUH. Full chimerism and increasing activity weredocumented after the second CBT with a follow-up period of 5 months by June2000.

The preliminary data show that the stored cord-blood units are suitable forclinical use.

60 Fourth International Symposium on Hematopoietic Stem Cell Transplantation

THE CURRENT STATUS OF UC BLOOD BANK IN KOREAH Han, The Catholic Umbilical Cord Blood Bank, College of Medicine, TheCatholic University, Seoul, KoreaSince 1996 when the first UC blood bank (UCBB) was established in Korea, threeUCBBs have been established and about 2350 UC blood (UCB) units have beenstocked. The quantities of UCB units stored in each UCBBs are as follows: theCatholic UCBB established in 1996 has 2000 units cryopreserved; Samsung(established in 1997) 150 units; and Yonsei (in 1997) 200 units. Each of the UCBBhas its own criterion for collecting and processing UCB, so the basic checklists forUCB banking are different for each centre. For this reason, an overall qualityassurance system for UCB units is required. Recently, the Catholic UCBBreported the results of the analysis of 1000 UCB units. The mean volume ofcollected UCB units was 82.7? and the average volume through the reductionprocess was 23.2?, which were stored in 2196?. The numbers of MNC(mononuclear cells) per UCB unit were in the range of 5.0–26.1 3 108 cells and themean MNC was 8.81 3 108 cells. After thawing of UCB, viability of MNC wasshown to be > 83.4%. The CD34+ cell per UCB units was assessed as 2.7%. Thefraction of CD34+CD38- cells was in the range of 0.3–3.3% and the median valuewas 0.5%. Out of 1000 UCB units, 10 units were contaminated. Through thisanalysis, the quality of UCB units cryopreserved in the Catholic UCBB wasconfirmed to be no different from that of foreign countries. The Catholic UCBBhas carried out 12 unrelated UCB transplantations for various hematologicdiseases, such as AML, CML and Gaucher’s disease. Successful engraftment wasidentified in nine patients. At present, two cases of unrelated UCB transplantationare performed monthly. In line with increasing the use of UCB units, 250 cases ofUCB transplantation twice every year are anticipated in the near future. It istherefore estimated that at least 10 000 UCB units have to be secured in Korea tocover domestic requirements. But I am sure this will not be sufficient to coverpatients. In conclusion, I feel that we Asians should cooperate to secure andestablish the network of UCBB with each other, for the health and welfare ofhumanity.

AUTOLOGOUS PBSC TRANSPLANTATION WITHOUT CRYO-PRESERVATIONT van Be, T van Binh, N tan Binh, Blood transfusion and Hematology Center,School of Medicine and Pharmacy, HCM City, VietNamIn most instances, patients who receive high-dose chemotherapy are rescued withstem cells that have been cryopreserved at -196°C. This is a complex andexpensive method. Between November 1996 and October 1998, we performed 15autologous PBSC transplantations without cryopreservation, patients werediagnosed with AML in first complete remission (CR1, n = 9), ALL in CR1 (n =2), CML in chronic phase (n = 3) and NHL in CR1 (n = 1). The median age ofpatients was 35 years (range 18–46) . PBSC mobilizing regimens consisted ofdaunorubicin (50 mg/m2/day, Days 1–5), cytarabine (400 mg/m2/day, Days 1–3)and etoposide (150 mg/m2, Days 1–3) in three CML patients, andcyclophosphamide (2 g/m2/day, Days 1–2) in 12 other patients (nine AML, twoALL, one NHL) followed by 300 µg of G-CSF daily, until the day before the lastleukapheresis. When a WBCC reached 4 3 10.9/L, leukapheresis was started andcontinued for 3 consecutive days. As soon as the third collection was finished,melphalan (180 mg/m2) was given as a conditioning regimen. Forty two hours aftergiving melphalan, the collected PBSCs that had been preserved at +4°C (90 h forthe first, 66 h for the second, and 42 h for the third collection) were reinfused. GM-CSF (300 µg/day) was given during the nadir period, until the absolute neutrophilcount (ANC) >1 3 109/L for 3 consecutive days. The median number ofmononucleated cells and CD34+ cells was 5.3 3 10.8/L (range 1.6–10.0 3108/L)and 6.3 x 106/L (range 0.5–19.1 3 106/L) respectively . All patients engrafted, withthe median time to recover ANC > 0.5 3 109/L and platelet count > 20.0 3 109/Lbeing 13 days (range 9–25 days) and 22 days (range 9–120 days) respectively.Following transplantation, four patients (three AML, one ALL) relapsed, 11patients (73.3%) remain alive and in CR with a median follow-up of 28 months(range 12–36 months). In conclusion, autologous PBSC transplantation withoutcryopreservation is a simple and economic method, but retains rapid recovery ofneutrophils and platelets, with stable engraftment .

MESSAGE TO THE SYMPOSIUM S Issaragrisil, Mahidol University, Bangkok, ThailandCord-blood has been used as an alternative source of stem cells for transplant. Inrelated cord-blood transplant, the results are comparable to those with othersources of stem cells. However, the incidence of GvHD is low. Our experiencewith sibling cord-blood transplant in thalassemia indicates favorable results; so far,< 14 cases have been successfully transplanted.

Only 25–30% of eligible patients can undergo sibling stem-cell processing,IMSUT transplantation. The rest do not have HLA-matched siblings as donors. ABM donor registry has been established in several countries in Asia, such as Japan,Korea, Taiwan, etc. In Thailand, we have not yet established one. We thereforewould like to develop cord-blood stem cell banking instead and are nowestablishing a cord-blood bank in Thailand. We badly need to organize the AsianCord Blood Network, in order to share cord-blood specimens to be used fortransplant. I personally would like to support the idea of setting up the Asian CordBlood Network. If I can do anything to help, I will be more than happy toparticipate.