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[CANCER RESEARCH 45, 4963-4969, October 1985]

Evaluation of Growth and Histology of Human Tumor Xenografts Implantedunder the Renal Capsule of Immunocompetent and Immunodeficient Mice1

James A. Bennett,2 Vernon A. Pilon, and Richard T. MacDowell

Departments of Surgery [J. A. B., R. T. M.] and Pathology [V. A. P.], Albany Medical College, Albany, New York 12208

ABSTRACT

Fresh surgical expiants of human carcinomas were implantedas first transplant generation xenografts under the kidney capsule of mice. Immunocompetent and immune-deficient mice were

compared for their ability to support the persistence and growthof these xenografts. Consistent growth of tumor xenograftscould not be demonstrated following implantation under thekidney capsule of immunocompetent mice. Immunological infiltration and rejection of the xenografts began 3 days postimplantation, and tumors were largely eliminated from the subcapsularspace by 6 days postimplantation. In contrast human tumorsconsistently grew under the kidney capsule of nude mice. Significant growth became apparent by 9 days postimplantationwith most human carcinomas and continued thereafter. Growthwas always accompanied by neovascularization of tumor xenografts which was visible by examination of tumor-bearing kidneys

under a dissecting microscope (x 6). There was no histologicalevidence of immunological interference with the persistence andgrowth of xenografts in nude mice. Thymectomized, irradiated,bone marrow-reconstituted conventional mice, as well as con

ventional mice, treated daily with 60 mg of cyclosporine A/kgwere comparable to nude mice as hosts which supported thelong-term persistence and growth of subrenal capsule implants

of human tumors. Such mice could provide an alternative tonude mice as hosts in which chemosensitivity assays could becarried out against growing human tumors at a considerablesaving in cost and convenience.

INTRODUCTION

The SRC3 is being used as an in vivo test of human tumorresponsiveness to drug therapy (1-7). Pieces (1 mm3) of solid

tumor are implanted as xenografts under the kidney capsule ofmice. Tumor-bearing mice are treated with a particular cancer

chemotherapeutic agent, and change in tumor size is recordedas a reflection of tumor sensitivity to that agent. Bogden ef a/.(1) developed the SRC in congenially athymic (nude) mice because of their inability to mount a host versus graft reactionagainst the human tumor xenograft. These investigators havesince adapted this assay to normal, immunocompetent micebecause of the high cost of nude mice (8). They reported thatthe host versus graft response in the subrenal capsule environment of immunocompetent mice did not significantly inhibit xenograft persistence during the first week after grafting. This

1Supported by the Elsa U. Pardee Foundation, the Fannie E. Rippel Foundation,American Cancer Society Grant IN-151 A, and National Cancer Institute Grant 1R01CA38033-01.

2To whom requests for reprints should be addressed.3The abbreviations used are: SRC, subrenal capsule assay; TIB, thymectc-

mized, irradiated, bone marrow-reconstituted.Received 1/23/85; revised 6/10/85; accepted 6/19/85.

conclusion was based on the observation of similar and in somecases greater size increases in human tumor xenografts innormal, immunocompetent mice compared to that found in nudemice in a 6-day assay (8). Several investigators have begun using

this assay to test the chemosensitivity of human and experimental tumors xenografted into the subrenal capsule space of eithernude mice (2) or immunocompetent mice (3-7, 9, 10).

Recent histological studies by Edelstein et ai. (3), Bennett etai. (11), Dumont ef al. (6), and Levi ef a/. (5) have demonstratedsignificant host cell infiltration of human tumor xenografts 6 daysafter implantation under the renal capsule of immunocompetentmice, and have suggested that this infiltration could contributeto size changes in the xenografts which could complicate interpretation of chemosensitivity data obtained in the assay. Thepresent study was designed to compare immunocompetent miceto nude mice as hosts for human tumor xenografts implantedunder the kidney capsule. The persistence and growth of thesexenografts, as well as their induction of histopathologicalchanges in the subrenal capsule environment during the first 3weeks after implantation, are described. Regimens of immuno-

suppressing conventional mice which could make possible thesubstitution of immunosuppressed conventional mice for nudemice as hosts for human tumor xenografts are also described.

MATERIALS AND METHODS

Mice

Male C57BL x DBA/2 F, (hereafter called BD2F,) mice were purchased from The Jackson Laboratory (Bar Harbor, ME) when they were5 weeks old. Male Tac:N[NIH] Swiss nu/nu (nude) mice were purchasedfrom Taconic Farms (Germantown, NY) when they were 5 weeks old.Nude mice were housed under positive laminar flow and handled usingsterile technique. Mice were not used in experiments until they were 7to 10 weeks old.

Immunosuppressive Regimens

Cyclosporine A. Cyclosporine A (Sandimmune i.v.) was purchasedfrom Sandoz, Inc., East Hanover, NJ and was diluted in 0.9% NaCIsolution. It was injected s.c. into BD2F! mice at a dose of 60 mg/kgevery day for the duration of the experiment.

Thymectomy, Irradiation, and Bone Marrow Reconstitution. T-cell-deprived mice were created by thymectomy followed by lethal irradiationand bone marrow reconstitution. Normal 5-week-old BD2F, mice were

thymectomized (12). Mortality rate from thymectomy was less than 10%.Fourteen days after thymectomy, mice were lethally irradiated (750 rads)and then reconstituted with 2 x 107 syngeneic bone marrow cells given

i.v. 24 h after irradiation. Bone marrow cells were pretreated withmonoclonal anti-Thy-1.2 (Becton Dickinson) plus complement. TIB mice

were used in experiments 1 to 3 weeks after bone marrow reconstitution.

Procurement of Human Tumor Specimens

Freshly resected human tumors were sectioned in the surgical pathology laboratory adjacent to the operating room suite under the guid-

CANCER RESEARCH VOL. 45 OCTOBER 1985

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GROWTH OF HUMAN TUMORS UNDER RENAL CAPSULE OF MICE

ance of a surgical pathologist in order to select for tissue which containeda maximum of viable tumor cells and a minimum of necrotic tissue,stroma, and nonneoplastic cells. Frozen sections were examined microscopically to ensure an adequate yield of tumor cells in the piece selectedfor the SRC. Once the tumor piece had been selected, it was placed ina sterile specimen container, immersed in Hanks' balanced salt solution

containing penicillin (100 units/ml) and streptomycin (100 M9/ml), andbrought to the laboratory for further dissection and implantation underthe kidney capsule of mice.

Implantation of Human Tumors under the Kidney Capsule of Mice

The method of implanting human tumors under the kidney capsule ofmice has been described by Bogden et al. (1 , 8). Briefly, mice wereanesthetized with Nembutal. The mouse kidney was exteriorized surgically, an incision was made in the capsule, and a 1-mm3 tumor fragment

was implanted under the capsule using a 16-gauge x 1.5-in. trocar. The

size of the implant was determined in situ by measuring two perpendicular diameters to the nearest 0.1 mm using a dissecting microscopeequipped with an ocular micrometer. The wound was closed with silkand wound clips, and each individual mouse was marked for identification. At various times after implantation, mice were sacrificed, the tumor-

bearing kidney was exteriorized, a final measurement of xenograft sizewas taken, and the xenograft-bearing kidney was resected and placed

in formalin. Change in xenograft size (AXS) was calculated using theformula:

AXS = W L + Wâ€”day n -- - â€” day 0

where L and IV represent two perpendicular diameters of the xenograft(8).

Histology

Preparation of slides for histology was carried out by cutting a 3-mm-thick cross-section containing the tumor tissue perpendicular to the long

axis of each kidney. Each section was embedded in paraffin, seriallysectioned by microtome, mounted on glass slides, and stained withhematoxylin and eosin. Histological studies provided qualitative resultsindicating that xenografts were occupied by varying amounts of tumorat various times after implantation. The following formula was utilized toapproximate the portion of the xenograft (X) occupied by tumor (T):

T/X% =c x d

x 100

The linear dimensions a, b, c, and d are defined in Chart 1. Chart 1 is asketch of a photomicrograph of a human adenocarcinoma of the colon6 days after implantation under the kidney capsule of Â¡mmunocompetentmice.

RESULTS

Fresh surgical expiants from two human adenocarcinomas ofthe colon (Chart 2, A and B) and one small cell carcinoma of thelung (Chart 2C) were implanted under the kidney capsule inimmunocompetent and athymic nude mice. The growth of thesehuman tumor xenografts in their respective hosts was comparedat 3-day intervals. As shown in Chart 2, no consistent increasein xenograft size occurred when tumors were implanted in immunocompetent mice. In contrast, in athymic nude mice a significant increase in xenograft size occurred by day 9 after implantation, and further increases in xenograft size were seen ondays 12, 15, and 18 after implantation. Moreover neovasculari-

zation, a marked increase in depth, and clear margins occurredin the xenografts growing in nude mice during the second and

Adenocarcinomaof ColonKidneyCapsuleImmuneInfiltrateandotherReactiveTissue

NormalKidneyTissue

Chart 1. Sketch of a histological section of a human adenocarcinoma xenograft6 days after implantation under the kidney capsule of an immunocompetent mouse.The dimensions of the tumor are estimated by the area encompassed by a x b.The dimensions of the implant space are estimated by the area encompassed byc xd. For comparison, see Fig. 2c, which is a photomicrograph of a histologicalsection of a human colon adenocarcinoma xenograft 6 days after implantationunder the kidney capsule of an immunocompetent mouse.

a x b% of tumor in zenograft = x 100

DAYS AFTER XENOGRAFT IMPLANTATION

Chart 2. Pieces (1 mm3) from two freshly resected human adenocarcinomas of

the colon (A and B), and one small cell carcinoma of the lung (C) were implantedas first transplant generation xenografts under the kidney capsule of normal (â€¢)and nude (O) mice. At various times after implantation mice were sacrificed, thetumor-bearing kidney was exteriorized, and xenograft size was measured grossly

under a dissecting microscope equipped with an ocular micrometer. Change inxenograft size was calculated as the difference between xenograft size measuredon the implant day and the assay day in the same mouse. Each value representsthe mean change in xenograft size Â±the range evaluated in two (A and B) or four(C) replicate mice.

third weeks after implantation (Fig. 1). These gross characteristics were never seen in the xenografts implanted in the immunocompetent mice. In fact the margins of the xenograft becamevery undefined and hazy during the second week after implantation in immunocompetent mice, making it difficult to measurexenograft size.

To assess the role of the host versus xenograft immuneresponse in preventing tumor growth in immunocompetent mice,we examined the histopathological changes induced by thexenografts described in Chart 2B in immunocompetent mice andcompared them to the changes induced by the same tumor innude mice. As shown in Fig. 2 (Fig. 2, a, c, e, and g) inimmunocompetent mice, infiltration of the tumor with mononu-

clear cells and neutrophils began on day 3, became very heavyon day 6, and resulted in complete rejection of the tumor by day9 after implantation. Tumor cells were never evident under thekidney capsule of immunocompetent mice during the secondand third weeks after implantation. Mononuclear cells, fibro-


4964




blasts, and collagen had replaced tumor during this interval.Furthermore, this event of tumor rejection and laying down ofscar tissue in immunocompetent mice was not accompanied bya consistent increase or decrease in xenograft size, and thuscould not be determined by macroscopic measurement alone(Chart 2). In contrast in nude mice there was no infiltration oftumor by mononuclear cells or neutrophils on day 3, someinfiltration of the tumor by mononuclear cells occurred on day 6,but there was no evidence of tumor rejection on day 9 afterimplantation (Fig. 2, b, d, f, and h). Tumor expanded in thesubrenal capsule space of nude mice during the second andthird weeks after implantation, and tumor cells undergoing mitosis were frequently seen during this interval (Fig. 2h).

Several other fresh surgical expiants of human tumors wereimplanted as first transplant generation xenografts under thekidney capsule of normal and nude mice and were examined 6days after implantation for amount of tumor in the xenograft andchange in xenograft size. The results in Table 1 indicate thatthere was no correlation between amount of tumor in the xenograft and change in xenograft size during the first 6 days afterimplantation. Decreases as well as increases in xenograft sizewere found where the major portion of the subcapsular implantspace was occupied by viable tumor. Changes in xenograft sizewere quite small, the largest being an increase of 0.4 mm inaverage diameter of the xenograft. In all cases the amount oftumor in the xenograft was markedly less in immunocompetentmice than it was in nude mice. In immunocompetent mice therewas significant immunological infiltration around the tumor andevidence of tumor necrosis in all ten cases. In five of ten casestumor had been completely eliminated from the subcapsularspace in immunocompetent mice. In contrast in nude mice the

Table 1Human tumor persistence and xenograft size changes in 6-day xenografts

implanted under the kidney capsule of normal and nude micePieces (1 mm3) of freshly resected human tumors were implanted as first

transplant generation xenografts under the kidney capsule of normal and nudemice. Each tumor was implanted into four mice, two normal and two nude. Sixdays after implantation mice were sacrificed and the xenograft-bearing kidney wasremoved and placed in 10% buffered formalin. Change in xenograft size (AXS) andthe percentage of tumor in the xenograft (T/X%) were calculated as described in"Materials and Methods." Each value represents the mean Â±the range from two

mice.

NormalPatient12345678910TumorAdenocarcinomaof

colonAdenocarcinomaof






colonSquamouscarci

nomaofesophagusAdenocarcinomaof

lungSquamouscarci

noma ofheadandneckT/X%13Â±900020

Â±406Â±62

+2025

Â±10AXS

(mm)+0.1

Â±0.2-0.02

Â±0.07+0.07

Â±0.08+0.05

Â±0.2+0.37

Â±0.07+0.42

Â±0.02-0.1

Â±0.05+0.1+0.4

Â±0.10Â±0.1T/X%50

Â±710062

Â±948

Â±1810072

Â±1210065

Â±170

Â±11100NudeAXS

(mm)+0.45

Â±0.05-0.1

Â±0.1-0.02

Â±0.02+0.2

+0.2+0.13

+0.07+0.22

Â±0.160

Â±0.05+0.17

Â±0.08-0.05

Â±0.1+0.22

Â±0.02

major portion of the implant site was occupied by viable tumorcells, there was no evidence of tumor necrosis, and there wasonly sparse infiltration by mononuclear cells in ten of ten cases.

The amount of tumor in human colon cancer xenografts wascompared to the macroscopic measurement of change in xenograft size at various times following implantation of tumor piecesunder the kidney capsule of normal mice, nude mice, and miceimmunosuppressed by alternative means, namely, (a) thymec-

tomy followed by lethal irradiation and bone marrow reconstitution (TIB mice), or (b) daily treatment with cyclosporine (60 mg/kg/day) (Table 2). As described above, tumor was largely eliminated from the subrenal capsule space of immunocompetentmice by day 6 after implantation. In contrast the major portion ofthe implant site was occupied by tumor in nude mice, TIB mice,and cyclosporine-treated mice at day 6. During the second week

after implantation tumor was completely eliminated from thesubrenal capsule implant site in immunocompetent mice. However, tumor persisted and grew within the subcapsular space ofimmunosuppressed mice. TIB and cyclosporine-treated micewere comparable to nude mice in supporting the long-term

persistence and growth of human tumor xenografts (Table 2). Infact in a separate series of studies which will be reportedelsewhere,4 human tumor was still present and expanding in the

subcapsular space of cyclosporine-treated mice 35 days after

implantation.

DISCUSSION

The results of this study indicated that human tumors implanted as xenografts under the kidney capsule of immunocompetent mice induced significant infiltration of murine host defenseelements into the subcapsular space, which resulted in immu-nological rejection of the tumor during the first week after implantation. This has been a consistent finding in over 12 tumorswhich we have studied and has not been due to a problem intissue selection, since the same tumors thrived with no evidenceof immunological rejection when they were implanted under thekidney capsule of nude mice. These results are in agreementwith the studies of Edelstein ef al. (3). However, recent studiesby Aamdal ef al. (7) have described significant growth of humantumor xenografts under the kidney capsules of both immunocompetent mice and nude mice in 6 days, leading these investigators to advocate use of immunocompetent mice in place ofnude mice and a 6-day assay time frame for tumor chemosen-

sitivity testing. The discrepancy between these results and thosedescribed both in this report and by Edelstein ef a/. (3) are mostlikely due to the fact that the studies carried out by Aamdal efal. (7) utilized fast-growing serially transplanted human tumor cell

lines rather than freshly resected first transplant generationhuman tumors. We have found significant growth of fast-growingtumor cell lines by day 4 after implantation under the kidneycapsule of immunocompetent mice, and have evaluated che-mosensitivity within this time frame (13). However, these tumorswere also heavily infiltrated with host immune cells by day 6 afterimplantation, preventing further xenograft growth. In our experience all cases of freshly resected human tumors implanted asfirst transplant generation xenografts have been slow growing

4J. A. Bennett, V. Pilon, D. R. Briggs, and M. F. McKneally. Evaluation ofcyclosporine-treatedmice as hosts for growing and testing the chemosensitivityoffirst transplant generation human tumor xenografts implanted under the kidneycapsule, submitted for publication, 1985.


4965




Table 2Persistence and growth of human colon cancer xenografts under the renal capsule of immunocompetent and immunodeficient mice

Pieces (1 mm3) of freshly resected human adenocarcinoma of the colon were implanted as first transplant generation xenografts under the renal capsule ofimmunocompetentand immunosuppressedmice. Immunosuppressedmice consisted of athymic nude mice; thymectomized, irradiated,bone-marrow-reconstitutedmice-and mice treated with cyclosporine (60 mg/kg/day s.c.) beginning on the day of implantation. At the indicated times after implantation,mice were sacrificed, the tumor-bearing kidney was exteriorized, and xenograft size was measuredgrossly under a dissecting microscope equipped with an ocular micrometer.Changein xenograft size(AXS) and percentage of tumor in the xenograft (T/X%) were calculated as described in "Materials and Methods."

Days afterimplantation3

69

1215NormalT/X%50

Â±37Â±7

000AXS

(mm)+0.1

Â±0.07+0.1 Â±0.2-0.2 Â±0.1+0.1 Â±0.03+0.1 Â±0.07T/X%50

Â±1358 Â±768 Â±2190 Â±4

100NudeAXS

(mm)0Â±0.15

+0.45 Â±0.05+0.7 Â±0.25+1.0 + 0.5+1.8 Â±0.7T/X%100

85 Â±563 Â±11

100ND"TIBAXS

(mm)+0.2

Â±0.070

+0.4+0.6 Â±0.2

NDCyclosporineT/X%75

Â±15100

65 Â±25100

90 Â±10AXS

(mm)0

-0.1 Â±0.05+0.3 Â±0.2+0.8 Â±0.4+1.5 Â±0.4

* ND, not determined

compared to tumor cell lines, and have been immunologicallyrejected in immunocompetent mice before the development ofsignificant tumor growth.

Other investigators have used immunocompetent mice ashosts for subrenal capsule implants of human tumors as a modelfor testing the chemosensitivity of these tumors during the firstweek after implantation (4-6, 9). In such a model the target of a

chemotherapeutic agent could be either the human tumor component, the murine host defense component, or both. Drugtoxicity to either of these components could result in reductionin xenograft size, which would complicate interpretation of tumorchemosensitivity. Levi et al. (5) have attempted to circumventthis problem by assaying 4 days rather than 6 days after implantation, and by evaluating each tumor-bearing kidney histologi-cally. Although there is less Â¡mmunological infiltrate in a 4-day

assay, in this model there is no way of knowing the amount oftumor in each xenograft until the assay has been completed andthe tumor-bearing kidneys have been examined histologically.

Not knowing the amount or the integrity of tumor in the xenograftbefore drug treatment could complicate interpretation of tumorchemosensitivity in this model. Also, under the best of circumstances slow-growing human tumors would persist but not necessarily grow in a 4-day assay, and 4 days may not be long

enough to observe the cytoreductive effects of certain drugsagainst such tumors.

Substitution of nude mice for immunocompetent mice eliminated infiltrating murine host defense components responsiblefor xenograft rejection. However, even in nude mice, we foundthat in most cases there were elements in addition to tumormaking up the xenograft. Examples of such elements werehuman stroma, phenotypic products from the tumor such asmucin, vascular tissue, and some murine lymphoreticular cellsand stroma. These contaminating noncancerous elements wouldcomplicate interpretation of chemosensitivity in a short-term

assay and underscore the need for careful selection of tumortissue for implantation. We have found histological examinationof frozen sections from the tumor to be implanted very helpful inselecting areas of the tumor which were highly enriched in viabletumor cells. We have also found that allowing the tumor to growfor 2 weeks under the kidney capsule of immunodeficient miceresulted in a marked expansion in the amount of tumor withinthe subcapsular space, which significantly reduced contaminating noncancerous tissue.

Persistence and growth of human tumors under the kidneycapsule of immunodeficient mice for 2 weeks provides a modelin which the chemosensitivity of human tumors can be tested in

vivo in a pharmacologically active system against growing solidtumors. Growth can be verified by examining the tumor-bearingkidney approximately 2 weeks after implantation during a second-look laparotomy. The mice tolerate this second surgical

procedure quite well with less than 5% mortality. The criteriawhich we have established to grossly identify functioning andgrowing tumor under the kidney capsule during this second-lookoperation are, (a) increase in xenograft breadth, (o) increase inxenograft depth, (c) neovascularization of the xenograft, and (d)clear xenograft margins (Fig. 1). Histological evidence of tumorhas been found in 100% of the cases in which these criteria havebeen satisfied, and these criteria have been satisfied in approximately 80% of the immunodeficient mice implanted with tumor.The remaining 20% presumably received noncancerous tissue,cancerous tissue which was not viable, or cancerous tissuewhich would not grow in the subrenal capsule space. The tumortake rate has been considerably lower than 80% in necrotictumors or in tumors which were grossly interspersed with non-

neoplastic tissue.Nude mice were not optimal as immunoincompetent hosts for

a subrenal capsule chemosensitivity assay, because they werecostly, not always available in large numbers from suppliers, andfastidious in their care and housing requirements. The results ofthis study indicated that the TIB or cyclosporine-treated conventional mice supported long-term persistence and growth of hu

man tumor xenografts under the kidney capsule, and could beused as an alternative to nude mice in a long-term subrenal

capsule chemosensitivity assay, providing a considerable savingin cost. Steel ef al. (14) have carried out extensive tumor chemosensitivity studies in TIB mice bearing s.c. implants of humantumors. These mice tolerated chemotherapy, but tumor take ratewas reduced when compared to take rate in studies wheretumors were implanted in the subrenal capsule space of immunosuppressed mice. We have tested the chemosensitivity ofcarcinoma xenografts growing in the subrenal capsule space ofcyclosporine-treated mice and have found the tumor chemosensitivity to be similar to that obtained in nude mice.4

The subrenal capsule space of mice is a suitable environmentfor growing and studying human solid tumors. The rich vascularbed in that environment bathes the tumor in nutrients, resultingin high tumor take rates (1, 8). Limiting factors for growingtumors in this environment such as host versus xenograft response and proper selection of tumor tissue can be overcome,respectively, by using appropriately immunosuppressed miceand by examining frozen sections of tumor tissue for guidanceto areas of the tumor rich in viable tumor cells. Previous studies


4966




by our group (13, 15) and by Maenpaa ef al. (16) have demonstrated accurate prediction of chemosensitivity against fast-

growing experimental solid tumors using this assay. However,the usefulness of this assay for predicting chemosensitivity ofhuman tumors remains to be determined. Pharmacological questions pertaining to timing the initiation and duration of drugtreatment and to measuring the duration of drug effect againstgrowing tumors still need to be worked out. The biologicalproblem of tumor heterogeneity which could potentially lead toa lack in uniformity of drug effect across a tumor specimen needsto be assessed. In spite of the need for further development,carrying out the SRC in immunosuppressed mice over a 3-week

time period stands as a unique in vivo model in which humansolid tumors can be reliably grown in a relatively short period oftime, and in which chemosensitivity testing can be carried outagainst tumors whose growth has been verified before drugexposure.

ACKNOWLEDGMENTS

We wish to thank Deborah R. Briggs for excellent technical assistance, andMancia E. Propp and Ruth D. Myer for their editorial and secretarial assistance inpreparation of the manuscript.

REFERENCES

1. Bogden, A. E., Kelton, D. E., Cobb, W. R., and Esber, H. J. A rapid screeningmethod for testing chemotherapeuticagents against human tumor xenografts.In: D. Houchens and A. Ovejera (eds.), Proceedings, Symposium on the Useof Athymic (Nude)Mice in Cancer Research,pp. 231-250. New York: GustavFischer, 1978.

2. Giovanella,B. C., Stehlin, J. S., Shepard, R. C., and Williams, L. J. Correlationbetween response to chemotherapy of human tumors In patients and in nudemice. Cancer (Phila.),52:1146-1152,1983.

3. Edelstein,M. Bâ€žFiebig, H. H., Smink, T., Van Putten, L. M., and Schuchhardt,C. Comparison between macroscopic and microscopic evaluation of tumor

responsivenessusing the subrenalcapsule assay. Eur. J. Cancer Clin. Oncol.,Ã•9:995-1009,1983.

4. Griffin, T. W., Bogden, A. E., Reich, S. D., Antonelli, D., Hunter, R. E., Ward,A., Yu, D. T., Greene, H. L., and Costanza, M. E. Initial clinical trials of thesubrenal capsule assay as a predictor of tumor response to chemotherapy.Cancer (Phila.),52: 2185-2192,1983.

5. Levi, F. A., Blum, J. P., Lemaigre,G., Bourut, C., Reinberg,A., and Mathe, G.A four-day subrenal capsule assay for testing the effectiveness of anticancerdrugs against human tumors. Cancer Res., 44: 2660-2667,1984.

6. Dumont, P., van der Esch, E. P., Jabri, M., Lejeune, F., and Atassi, G.Chemosensitivity of human melanoma xenografts in immunocompetent miceand its histologicalevaluation. Int. J. Cancer, 33: 447-451,1984.

7. Aamdal, S., Fodstad, O., and Pini, A. Human tumor xenografts transplantedunder the renal capsule of conventional mice. Growth rates and host immuneresponse. Int. J. Cancer, 34: 725-730,1984.

8. Bogden, A. E., Haskell, P. M., LePage, D. L., Kelton, D. E., Cobb, W. R., andEsber, H. J. Growth of human tumor xenografts implanted under the renalcapsuleof normal immunocompetentmice. Exp. Cell Btol.,47:281 -293,1979.

9. Bogden, A. E., Cobb, W. R., LePage,D. L., Haskell,P. M., Guikin,T. A., Ward,A., Kelton, D. E., and Esber, H. J. Chemotherapy responsivenessof humantumors as first transplant generation xenografts in the normal mouse. Cancer(Phila.),48: 10-20, 1981.

10. Raina, S., Hill, H. Z., Hill, G. J., and Rush, B. F. Scheduling of combinationchemotherapy for a murine melanoma with the subrenal capsule assay. J.Surg. Oncol., 26: 51-52,1984.

11. Bennett, J. A., Nguyen, M., Pilon, V., and MacDowell, R. T. Histopathologicalchanges induced by human colon cancer xenografts implanted under thekidney capsuleof immunocompetentand immunosuppressedmice. Proc. Am.Assoc. Cancer Res., 25: 372, 1984.

12. Ferguson,A. Thymectomy of the adult mouse. In: D. W. Weir (ed.),Handbookof Experimental Immunology, pp. A3.17-A3.18. Oxford: Blackwell ScientificPublications,1973.

13. Bennett, J. A., Pilon, V. A., Uppal, G. S., and McKneally, M. F. Accurateprediction of experimentalcancer chemosensitivityusing the subrenalcapsulexenograft assay. J. Surg. Oncol., in press, 1985.

14. Steel, G. G., Courtenay, V. D., and Peckham, M. J. The response to chemotherapy of a variety of human tumor xenografts. Br. J. Cancer, 47: 1-13,1983.

15. Bennett, J. A., McKneally,M. F., Prevosti, L., and Pilon,V. Accurate predictionof the chemosensitivity of Lewis lung carcinoma using a subrenal capsuleassay in rats. Proc. Am. Assoc. Cancer Res., 25: 373,1984.

16. Maenpaa, J., Gronroos, M., Leppanen, M., and Kangas, L. The subrenalcapsule assay in mice. Prediction of rat tumor sensitivity to chemotherapy.Cancer (Phila.),54:1530-1534,1984.

Fig. 1. Low power macroscopic picture of a human colon cancer xenograft 18 days after implantation under the kidney capsule of a nude mouse. Note well-definedmargins of xenograft and network of prominent capsular vessels providing vascular support for the tumor, x 12.


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1985;45:4963-4969. Cancer Res James A. Bennett, Vernon A. Pilon and Richard T. MacDowell Immunocompetent and Immunodeficient MiceXenografts Implanted under the Renal Capsule of Evaluation of Growth and Histology of Human Tumor

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