Results of the Cord Blood Transplantation Results of the Cord Blood Transplantation Study (COBLT) Unrelated Donor Banking Study (COBLT) Unrelated Donor Banking

Program from Donor Screening to Program from Donor Screening to Characterization of Banked UnitsCharacterization of Banked Units

Joanne Kurtzberg, MD 1, Elizabeth L Wagner, MPH 2, John K Fraser, PhD 3, Mitch S Cairo, MD 4, LeeAnn Jensen PhD 5, Geoff Cohen MS 2, Shelly L Carter, ScD 2, Nancy A Kernan, MD 6 and *** 7.

1 Duke University Medical Center, Durham, North Carolina, 27705 ; 2 The EMMES Corporation, Rockville, Maryland, 20850 ; 3 University of California, Los Angeles, California, 90095 ; 4 Georgetown University Medical Center, Washington DC, 20007 ; 5 National Heart, Lung and Blood Institute, Bethesda, Maryland, 20817 ; 6 Memorial Sloan-Kettering Cancer Center, New York, New York, 10021 and 7 ***sponsored by the National Heart, Lung and Blood Institute, Bethesda, Maryland, 20817

BackgroundBackground COBLT: The Cord Blood Transplantation COBLT: The Cord Blood Transplantation

Study – sponsored by the NHLBIStudy – sponsored by the NHLBI– Banking studyBanking study– Transplant StudyTransplant Study

Initiated in 1996 and funded:Initiated in 1996 and funded:– 1 Coordinating Center (EMMES)1 Coordinating Center (EMMES)– 3 Cord Blood Banks3 Cord Blood Banks

Choc/GTChoc/GT Mitch CairoMitch Cairo DukeDuke Joanne KurtzbergJoanne Kurtzberg UCLAUCLA John FraserJohn Fraser

– 7 Transplant Centers 7 Transplant Centers 21 additional centers added over tenure of the 21 additional centers added over tenure of the

studystudy

Banking Study: Plan of Banking Study: Plan of AttackAttack

Steering Committee: Steering Committee: – Chair, Nancy Kernan, MDChair, Nancy Kernan, MD– Bank DirectorsBank Directors– Coordinating & StatisticalCenter - EMMESCoordinating & StatisticalCenter - EMMES– Representatives from NHLBIRepresentatives from NHLBI– Transplanters, OB, Ethicist, Lab TechsTransplanters, OB, Ethicist, Lab Techs

Created and validated SOPs for Created and validated SOPs for programprogram– Monthly meeting x 18 monthsMonthly meeting x 18 months

Standard Operating Standard Operating ProceduresProcedures

SOPsSOPs– Donor screening and recruitmentDonor screening and recruitment– Donor education and consentDonor education and consent– Maternal donor medical history and Maternal donor medical history and

testingtesting– Collection and transportationCollection and transportation– Processing and cryopreservationProcessing and cryopreservation– Maternal Sample and Cord Blood TestingMaternal Sample and Cord Blood Testing– Quarantine and releaseQuarantine and release– Long term storageLong term storage– Search RegistrySearch Registry

SOPs - ContinuedSOPs - Continued

Standardization and validation of Standardization and validation of laboratory procedureslaboratory procedures– HLA by molecular techniquesHLA by molecular techniques– Cell counts, viability, CFUs, CD34 and Cell counts, viability, CFUs, CD34 and

subsets, CD3,4,8, CD19, CD16/56subsets, CD3,4,8, CD19, CD16/56 Creation of internet based banking Creation of internet based banking

data system and search registrydata system and search registry Interlaboratory/bank QA/QC program Interlaboratory/bank QA/QC program

with sample exchanges for testing, with sample exchanges for testing, HLA typing, banking and thawingHLA typing, banking and thawing

Study EligibilityStudy Eligibility Collections ex uteroCollections ex utero UCBs >40 ml or 6x10e8 TNCUCBs >40 ml or 6x10e8 TNC Initiation of processing within 48h of Initiation of processing within 48h of

collectioncollection Recovery of >60% of TNC post processingRecovery of >60% of TNC post processing

– +CFU growth+CFU growth Negative maternal ID testing and Negative maternal ID testing and

maternal and family risk historymaternal and family risk history Negative homozygous hemoglobinopathyNegative homozygous hemoglobinopathy All banked units passed a study algorithm All banked units passed a study algorithm

defined by the steering committeedefined by the steering committee

TimelineTimeline

Recruitment initiated in October, Recruitment initiated in October, 19971997

Collections initiated in December, Collections initiated in December, 19971997

SOPs published 1998SOPs published 1998– Fraser et al, J Hematotherapy Fraser et al, J Hematotherapy

7:521-561.7:521-561.– <www.emmes.com><www.emmes.com>

Collections completed June, 2001.Collections completed June, 2001.

Figure 1 - Numbers of Women Screened, Approached, Consenting and with Collections Made Data from Monthly

Recruitment Forms April 1998 to March 2001

Totalconsented

20710

Totalapproached

24240

TotalScreened

34799

Totalconsenting

and notexcluded

beforedelivery20278

Totalcollectionsrecorded on

monthlyrecruitment

forms

16764

Total not approached: 10559

Reasons for not approachingpotential donors

Pregnancy related 63%Personal medical history 12%Personal risk behavior 8%Language barrier 7%Family history 3%Unable to consent 3%Private cord blood bank 2%Other / not documented 2%

70% 98%85% 83%

30%

Total refusals3530

15%

Total excluded prior to delivery: 432

Reasons for excluding consenting

donors prior to delivery

Pregnancy related 20%Personal medical history 41%Personal risk behavior 20%Family history 9%Other 10%

2%

Total collection failures at delivery: 3514

Reasons for failure to collect

Collection staff not at center orunavailable 44%

Placenta/cord exclusion(including placental abruption) 41%

Other delivery exclusion 7%Miscellaneous/undocumented 8%

17%

Figure 2 - Numbers of Collections Made, Cryopreserved and Banked (December 1997 to August 2001)

Reasons for discard

Number PercentInsufficient volume/cell count 5171 64Positive maternal ID test 646 8Maternal history exclusion 613 8Processing issues 297 4Collection issues 294 4No maternal sample 243 3Subsequent medical information 224 3Failed post-processing sterility test 194 2Insufficient cell recovery 170 2Withdrew consent 90 1Administrative issues 57 1Shipping/storage issues 91 1Missing 5 0Total 8095 100

Cryopreserved11096

Totalcollections

17207

Moved to long-term storage9112

Already shipped 303Available in bank 8133Still in quarantine 676

82%64%

Discarded6111

36%

Discarded1984

18%

Table 1 - Influence of Type of Delivery on Cell Counts and Volume

Note : All differences highly significant (P< 0.0001).

VaginalVaginal C-sectionC-section ALLALL

N=6736N=6736 N=2315N=2315 N=9051N=9051

Pre-processingPre-processing

Volume (ml)Volume (ml) 76.676.6 89.289.2 79.879.8

Total viable nucleated cells (x10Total viable nucleated cells (x1088)) 11.611.6 12.212.2 11.811.8

Post-processingPost-processing

Total viable nucleated cells (x10Total viable nucleated cells (x1088)) 9.19.1 9.69.6 9.29.2

Total CD34+ cells (x10Total CD34+ cells (x1066) ) 3.33.3 3.63.6 3.43.4

Average CD34+cells per Average CD34+cells per uLuL 152152 169169 157157

Total CD3+ cells (x10Total CD3+ cells (x1088) ) 1.71.7 2.02.0 1.81.8

Average CD3+ cells per Average CD3+ cells per uLuL 80098009 90889088 82868286

Table 2 - Weight, Volume and Cell Counts of Stored CBUs by Ethnic Group

CaucasianCaucasianAfricanAfrican

AmericanAmericanAsianAsian HispanicHispanic

Mixed/Mixed/

OtherOtherAllAll

N=3701N=3701

40%40%

N=1316N=1316

14%14%

N=1114N=1114

12%12%

N=1910N=1910

21%21%

N=1071N=1071

12%12%

N=9112N=9112

Pre-processingPre-processing

Volume (ml)Volume (ml) 80.680.6 79.479.4 80.580.5 78.678.6 78.978.9 79.879.8

Total viable nucleated cells x10Total viable nucleated cells x108 8 12.312.3 10.910.9 11.411.4 11.711.7 11.611.6 11.811.8

Post-processing Post-processing

Total viable nucleated cell x10Total viable nucleated cell x1088 9.69.6 8.58.5 9.09.0 9.29.2 9.19.1 9.29.2

Total CD34+ cells x 10 Total CD34+ cells x 10 66 3.53.5 2.92.9 3.33.3 3.53.5 3.23.2 3.43.4

Average CD34+cells per Average CD34+cells per uLuL 163163 137137 153153 162162 151151 157157

Total CD3+ cells x 10Total CD3+ cells x 1088 1.91.9 1.61.6 1.81.8 1.81.8 1.71.7 1.81.8

Average CD3+ cells per Average CD3+ cells per uLuL 86258625 75197519 83698369 83698369 78767876 82928292

Table 3 - Percent of Collections Stored by Mother's Ethnicity

RaceRace All All CollectedCollected DiscardedDiscarded StoredStored Percent Percent

StoredStored

CaucasianCaucasian 6784 6784 (39%)(39%) 30833083 37013701 55 %55 %

African African AmericanAmerican

2842 2842 (17%)(17%) 15261526 13161316 46 %46 %

AsianAsian 19141914(11%)(11%) 800800 11141114 58 %58 %

HispanicHispanic 34943494(20%)(20%) 15841584 19101910 55 %55 %

MixedMixed 21732173(13%)(13%) 11021102 10711071 49 %49 %

TOTALTOTAL 1720717207 80958095 91129112 53 %53 %

Table 4 - Correlations of Cell Counts, CBU Volume, Infant Birth Weight and Gestational Age

*** P<0.001

VolumeVolume Birth Birth WeightWeight

Gestational Gestational AgeAge TNCCTNCC CD34CD34

Birth WeightBirth Weight 0.28***0.28***

Gestational Gestational AgeAge

0.000.00 0.34***0.34***

TNCCTNCC 0.66***0.66*** 0.25***0.25*** 0.12***0.12***

CD34CD34 0.31***0.31*** 0.12***0.12*** -0.04***-0.04*** 0.48***0.48***

CD3CD3 0.48***0.48*** 0.13***0.13*** 0.03***0.03*** 0.59***0.59*** 0.31***0.31***

Figure 2a - Total Nucleated Cells by Birth Weight

<2.5 2.5 - 3.0 3.0 - 3.5 3.5 - 4.0 > 4.0

7

8

9

10

11

12

TN

C (

x 10

8 )

Birth weight (kg)

<2.5 2.5 - 3.0 3.0 - 3.5 3.5 - 4.0 > 4.0

2.0

2.5

3.0

3.5

4.0

4.5

5.0C

D34

(x

106 )

Birth weight (kg)

Figure 2b - Total CD34 Cells by Birth Weight

Bars show mean and 95% confidence limits (mean +/- 1.96 standard errors).

<37 38 39 40 > 40

7

8

9

10

11

12

TN

C (

x 10

8 )

Gestational Age (weeks)

Figure 2c - Total Nucleated Cells by Gestational Age

Figure 2d - Total CD34 Cells by Gestational Age

<37 38 39 40 > 40

2.0

2.5

3.0

3.5

4.0

4.5

5.0C

D34

(x

106 )

Gestational Age (weeks)

Figure 3 - Adjusted Mean Total Post-ProcessingCD34 Counts by Race

Caucasian African

AmericanAsian Hispanic Other

1.5

2.0

2.5

3.0

Ad

just

ed m

ean

CD

34+

(x

106 )

To

tal

po

st p

roce

ssin

g c

ou

nt

ConclusionsConclusions

The COBLT Banking Study has The COBLT Banking Study has demonstrated the resources demonstrated the resources required to establish and required to establish and maintain a network of cord blood maintain a network of cord blood banks with common SOPs and banks with common SOPs and Quality Systems.Quality Systems.

A MCC (EMMES) created and A MCC (EMMES) created and maintains a web based data maintains a web based data management system for data management system for data collection and analysis.collection and analysis.

ConclusionsConclusions >35,000 women were recruited and >35,000 women were recruited and

screened over 3 years.screened over 3 years. 15% refused to participate in the study.15% refused to participate in the study. Approximately 53% of CBUs collected Approximately 53% of CBUs collected

from these donors were successfully from these donors were successfully banked creating an ethnically diverse banked creating an ethnically diverse inventory of >8,000 total units.inventory of >8,000 total units.

The main reasons for exclusion were The main reasons for exclusion were insufficient cell volume/counts or insufficient cell volume/counts or positive ID testing on the maternal positive ID testing on the maternal sample.sample.

ConclusionsConclusions Strong correlations were observed Strong correlations were observed

between UCB collection volume and between UCB collection volume and nucleated cell and CD34 content.nucleated cell and CD34 content.

Units collected from African American Units collected from African American donors had lower cell counts and donors had lower cell counts and CD34 counts per ml of UCB collected. CD34 counts per ml of UCB collected.

Factors correlating with higher UCB Factors correlating with higher UCB TNC content included higher birth TNC content included higher birth weight, delivery by c-section and weight, delivery by c-section and increasing gestational age at delivery increasing gestational age at delivery (not beyond 40 weeks). (not beyond 40 weeks).

ConclusionsConclusions

These observations can be These observations can be used to create an algorithm used to create an algorithm for identification of desirable for identification of desirable units at a collection site to units at a collection site to maximize resources maximize resources committed to units that are committed to units that are processed, tested and processed, tested and banked.banked.

AcknowledgementsAcknowledgements

NHLBI: LeeAnn Jensen, Paul McCurdyNHLBI: LeeAnn Jensen, Paul McCurdy Study Chair: Nancy KernanStudy Chair: Nancy Kernan The MCC – The EMMES CorporationThe MCC – The EMMES Corporation

– Shelly Carter, Liz Wagner, Geoff CohenShelly Carter, Liz Wagner, Geoff Cohen Bank DirectorsBank Directors

– Mitch Cairo, John Fraser, Joanne Mitch Cairo, John Fraser, Joanne KurtzbergKurtzberg

HLAHLA– UCSF, UCLA, NMRIUCSF, UCLA, NMRI