Transplantation of Human Tumors Into Cortisone-Treated Hamsters*

Transplantation of Human Tumors into Cortisone-treated Hamsters*

W. BRADFORDPATTERSON,!ROSANNAN.CHUTE,ANDSHELDONC.SOMMERSA

(Cancer Research Institute, New England Deaconess Hospital, Boston 15, Mass.)

The desirability of obtaining living human for animals weighing less than 75 gm. and 2 mg. malignant tissue for experimental purposes con for those over this weight, was given subcutane- tinues to grow in proportion to our increasing ously 3 times a week starting on the day of im knowledge of the genesis and treatment of cancer. plantation. Tumors were obtained fresh from the Although several methods for growing human operating room and transplanted as rapidly as cancer outside of the primary host have been de possible. The total time required for this process scribed and utilized (7-10, 13), human tumors varied from 30 minutes to 3 hours. Frozen sections which can be grown in predictable amounts, with were prepared in most instances, and care was in a reasonable time, and by different investigators taken to select fragments of tumor free of necrosis in laboratory animals, have not been reported. and gross infection. The tissue was not, as a rule, However, from the work of Toolan (14) it can be sterile on receipt, and no effort was made to steri surmised that after 70 years of effort the tools and lize it. However, sterile instruments were used, knowledge are at hand with which permanent fragments from within the received tissue were and useful heterotransplantation may be accom selected, and the recipient site was gently and plished. briefly cleansed with Zephiran (1:1000). The tis The purpose of this paper is to report a human sue was kept on a sponge wet with saline while epidermoid carcinoma, now established in cor fragments roughly 1-2 mm. in diameter were cut tisone-treated hamsters, which shows every evi and placed in No. 18 trocars. Transplantation dence of possessing sufficient "growth potential" was performed with the animal under intraperi- to continue as a permanently transplantable tu toneal nembutal anesthesia, depositing the tumor mor, growing with sufficient rapidity and certain near the tip of the everted cheek pouch. The ty to make it of value for experimental use. This pouch was handled as little as possible in order to tumor is one of a series of 37 human neoplasms minimize trauma and reaction. Animals having transplanted in the past 7 months. Histological tumor transplants were anesthetized every 5â€”7 sections are available in 33 of these cases, verify days for inspection of the pouches. Transplants ing "survival" in 23, or 70 per cent. were made to both cheek pouches. On a few oc casions, two or three fragments rather than one MATERIALS AND METHODS have been placed in each pouch. This has had no The hamster cheek pouch technic described by apparent effect on the growth or failure of the Lutz et al. (11) was selected for use because of its transplant. promise as a simple method with easily observa Transplants have been removed for study by ble results. Golden hamsters of both sexes, 3-5 everting the cheek pouch and excising part or all weeks old (40-70 gm.) were used. They were ob of the tumor. Sterile technic was used if the ani tained from a local professional breeder. A num mal was not to be sacrificed. Hemostasis was ob ber of older animals have been employed for par tained by brief local pressure. Healing of the mu ticular experiments. They were fed Purina Labo cosa is definitely retarded by treatment of the ratory Chow and water ad libitum, with lettuce hamster with cortisone, but a small mucosal rent provided once a week. Cortisone acetate, 1 mg. is well tolerated in an otherwise unaffected pouch. * This work was supported by funds from the American Partial removal of the transplant is often followed Cancer Society and United States Atomic Energy Commission by local infection, which may subside without Contract AT(SO-1)-901 with the New England Deaconess Hos treatment or become invasive. pital. t Fellow in Cancer Research of the American Cancer So A few transplants have been biopsied within ciety. Present address: Peter Bent Brigham Hospital, Boston, the 1st week, but those utilized in calculating tu Mass. mor survival statistics were all at least 12 days ÃŽPresent address : Massachusetts Memorial Hospitals, old. Specimens for histological examination were Boston, Mass. placed in Zenker's fixative and stained with Received for publication May 14, 1954. hematoxylin-eosin. 656

RESULTS repetitive, frequent transplantation. Two other Table 1 lists the results of this series. The usual tumors, to be discussed in a later paper, one epi- transplant shows a two- or threefold increase in dermoid and one embryonal carcinoma, also size in the 1st week, with slight increase in vas- promise to be permanently transplantable. "Deac-l."â€”This tumor originated in the pa cularity. The enlargement may be due to any one of several factors; there may clearly be growth of rotid gland of a 74-year-old man. The pathologi the tumor, with little host reaction except for an cal diagnosis was muco-epidermoid carcinoma, enlarging vascular supply; the existing tumor grade II (Fig. 3). It was implanted in both cheek cells, if glandular in function, may produce a pouches of six hamsters within 30 minutes of its sizable amount of secretion; a host reaction of removal from the patient, and four of these ani inflammatory exÃºdate, edema, and hemorrhage mals were treated in the standard fashion with may develop in spite of treatment with cortisone. TABLE1 We feel some confidence now in distinguishing these reactions grossly. Growing tumors invari SURVIVALOFHUMANTUMORTRANSPLANTSIN ably have a healthy pink color, shading to blue CORTISONE-TREATEDHAMSTERS according to their intrinsic vascularity. The Cases Positive* Negative borders are sharp, and there is no gross evidence Adenocarcinomas: 1 Parotid 0 of host reaction. Mucinous secretion has been Lung 1 recognizable as such in and around a few trans Breast o Pancreas 0 plants. On the other hand, the presence of a mud Sigmoid 0 dy and variegated coloring with indistinct borders Endometrium 0 indicative of edema is invariably associated with Prostate 0 a considerable host reaction. In such transplants Carcinoma simplex (breast) there may be a large portion of viable tumor cen Epidermoid carcinomas: 0 7 e Mouth 1 0 1 trally, but progressive growth does not occur. In Esophagus 1 1 o short, if the host can react to an appreciable ex Lung 3 3 0 tent, it usually wins. Grossly infected transplants Renal pelvis 1 1 I) Vulva 1 1 0 have dead-white or yellow areas indicating ab Penis 1 1 I) scess formation. Treatment with cortisone is ap Mixed tumor of parotid 2 â€¢i 0 parently responsible for elimination of the marked Undifferentiated (scalp) 1 \ 0 granulomatous reaction which in earlier experi Carcinoid (liver) 1 \ o Seminoma (testis) 1 0 1 ments (1) was difficult to distinguish from viable Small-cell carcinoma (lung) 1 0 1 tumor. Embryonal carcinoma (testis) 1 1 0 The growth period may last from 1 to 3 weeks, Fibrosarcoma (shoulder) 1 1 0 Lymphocytoma 1 I) 1 and in some cases is followed by partial or com Glioblastoma (brain) 1 0 1 plete regression during the ensuing days or weeks. Basal-cell carcinoma 1 1 0 A few transplants show initial growth, then sta Giant-cell tumor (spine) 1 1 0 bilization and persistence for long periods of time. Totals: 88 23 10 One carcinoma of the breast (Fig. 1) has recently * Histologie evidence of cell survival and/or growth for a period of at least lÃ¬days. Microscopic sections demonstrating questionable survival of been transplanted to a second generation of ham only a few cells were classed as negative. sters after surviving in the original animal for 116 days. This tumor showed many of the char cortisone. From the start the tumor showed ex acteristics of growth reported for anterior chamber ceptional growth, as evidenced by rapid and transplants (3, 6); that is, it grew initially for a marked increase in size and vascularity of five period of 4 weeks and then regressed. After an of the transplants. After 14 days in the first-gen interval of 6 weeks, during which time another eration animals, four tumors were large enough breast carcinoma was implanted and failed to to permit another transplantation. This process grow in the same pouch, the original nodule slowly has now been repeated to the 16th generation in enlarged tenfold. Biopsy at the time of the second the course of 9 months. In view of the marked transplantation showed a healthy, slowly growing increase in total mass of tumor now available, carcinoma (Fig. 2). Microscopically, it is difficult there seems little reason to anticipate a reversal to differentiate this from a tumor which shows of form and loss of the tumor. only "persistence," but in view of the substantial These transplants have an initial latent period increase of the gross tumor mass, proliferative of from 4 to 15 days, followed by an increase in growth has very obviously taken place. the vascular supply to the tumor and then steady The tumor to be described below is one of the enlargement. The tumor maintains a healthy series showing the striking growth which permits pink color, a firm consistency, and a well-vascu-

larized surface. No detailed studies on growth cortisone treatment. Most animals have tolerated rates have as yet been carried out, but in 3 weeks the regimen of 6 mg/week fairly well. A few ani the successful transplants of this tumor reach an mals have died with post mortem findings sug average diameter of 0.4-0.7 cm., and at 6 weeks gestive of congestive heart failure. Animals treated several tumors have weighed 0.3-0.5 gm. (A few for more than a month show generalized debility experiments have been done which indicate that, and increased deposition of intrascapular brown with multiple initial fragments, resultant tumors fat. are proportionately larger.) After the tumors have Experiments have been carried out on the use grown for 3-6 weeks, central liquefaction and de of hydrocortone (Compound F), locally, to avoid generation are often evident. In several tumors the systemic effects of cortisone. While this treat which have been allowed to remain for periods up ment will depress the host inflammatory response to 50 days, this degeneration has been followed and permit some tumor growth, it does not sup by ulcÃ©rationthrough the pouch, with the forma port growth adequately. tion of a lesion resembling many human carci Transplants of the Deac-1 tumor have been nomasâ€”a central ulcer with a rim of healthy tu made into 50 cortisone-treated hamsters. In this mor. Some of the transplants have regressed sub group, microscopic survival has been noted in 84 sequent to the infection and inflammation which pouches of 45 animals, and active growth in 80 have followed the ulcÃ©ration,but others are still of these. The tumor is at present growing in eight enlarging slowly at 60 days. No mÃ©tastaseshave een hamsters in this laboratory in varying stages. been observed, and no animal has died as a direct It is also being successfully grown and utilized in result of the tumor, although local recurrences three other laboratories in the Boston area. Trans have developed following what was regarded as plantation from one animal to the next has been an adequate excisional biopsy. accomplished at varying times between 10 and All transplants were verified microscopically. 52 days. Most transplants are made between 14 Sections of growing tumor showed a healthy and 21 daysâ€”when the tumors are sizable, and epidermoid carcinoma, grade II, with numerous degeneration is at a minimum. mitoses (Fig. 4). There was often evidence of local invasion, especially of the overlying mucosa. A DISCUSSION moderate amount of vascular stroma was present, The use of cortisone to eliminate the initial and host inflammatory response, if present at all, defense of an animal host to a foreign tumor, as included varying degrees of peripheral vasodilata first described by Toolan, has removed one of the tion, edema, and a scant lymphocytic infiltrate. most formidable obstacles to successful growth Infected specimens excited a severe inflammatory of tumors in animals. The epidermoid carcinoma exÃºdate in spite of cortisone, with abscess forma and fibrosarcoma reported by Toolan and the one tion and granulation tissue. Tumors that failed to reported here are vigorous tumors, selected for grow showed degeneration, with fibrosis, mum tuitously, which seem to possess sufficient growth mification, and calcification. There have been potential to cross the species barrier, with the aid few changes in the character of the neoplasm dur of cortisone, and grow progressively. In reviewing ing the 9 months. No mucoid element has been previous attempts to transplant tumors heter- seen since the first transplant, and a number of ologously, one is struck by the occasional wide recent transplants have shown an adeno-acan- variation in survival time reported by different thomatous character (Fig. 5). investigators and with different methods. This Three animals in recent generations have been is suggestive of a "spectrum" of tumors, which given no cortisone. Each implant has shown an may be transplanted permanently to animals one initial enlargement, with the edema and discolora by one as our knowledge increases. tion of inflammation, followed by rapid regression It is our conviction that there are many reasons within 10 days. Cortisone has also been stopped for failures with heterologous transplantation of on a number of animals bearing established trans malignant tumors. Spontaneous tumors in hu plants, with rapid rÃ©sorptionofthe tumor nodule. mans and heterologous transplants in animals Studies are in progress relative to the optimum both excite varying responses, indicative of mul-

Fio. 1.â€”Carcinoma simplex of breast. X 250. Fio. 4.â€”Deac-1, third-generation transplant in hamster. Pio. t.â€”Same tumor growing in hamster 116 days after X250. transplantation. X 250. FIG. 5.â€”Deac-1, ninth-generation transplant showing FIG. 3.â€”Muco-epidennoid carcinoma of parotid gland, the adeno-acanthomatous tendency, with hamster cheek pouch on Deac-1 tumor. X 250. right. X 125.

*\ ;;. .. **â€¢**'*^.Â« 5

Downloaded from cancerres.aacrjournals.org on September 25, 2021. © 1954 American Association for Cancer Research. PATTERSONetal.â€”Transplantation of Human Tumors into Hamsters 659 tiple lines of host defense. The stromal response SUMMARY determines the adequacy of the blood supply. 1. Transplantation of 33 human tumors into Inflammatory exÃºdate and allergic responses may the cheek pouch of cortisone-treated hamsters regulate survival and spread. Endocrine factors has resulted in the "survival" of 70 per cent of have been shown to be significant in the growth the tumors for periods exceeding 12 days. rate of both human and animal tumors. The nu 2. One epidermoid carcinoma, now called tritional state of the animal has long been known "Deac-1," holds promise of being permanently to be related to the survival of transplanted tu transplantable. It has now been carried through mors. Eichwald and others (5) have shown the sixteen generations of hamsters in a 9-month importance of a number of host and graft factors period. and feel there are many others. It seems reasona 3. Factors concerned in the eventual trans ble to compare our present-day technics for heter- plantation of other human neoplasms are briefly ologous transplantation with the initial attempts discussed, and possibilities for immediate clinical to culture bacteria. The most hardy varieties then application of experimental data are suggested. grew in the simplest media, but the culturing of more sensitive types of micro-organisms awaited REFERENCES the development of either more fortunate or more carefully controlled substrates. There are several 1. CHUTE,R. N.; SOMMEES,S.C.; and WABREN,S. HÃ©tÃ©ro- transplantation of Human Cancer. II. Hamster Cheek ways in which the less hardy tumors might be Pouch. Cancer Research, 12:912-14, 1952. encouraged to grow; for example, the anterior 2. DOBYNS,B.M., and LENNON,B.Changes in thÃ©Histologi- chamber transplants so intensively investigated cal Pattern of Human Thyroid Cancer after Serial Trans plantation in the Anterior Chamber of Eyes of Guinea by Greene circumvent the species barrier because Pigs. Cancer, 6:45-51,1952. the particular medium allows the transplant an 3. EICHWALD,E.J. Heterologous Transplantation of Cancer initial period of recovery and adjustment before of Childhood. Cancer Research, 8:273-77,1948. vascularization brings host defenses to bear (4). 4. EICHWALD,E.J., and CHANO,H. Y. The Significance of These transplants have certainly been successful the Anterior Chamber in Tumor Transplantation. Cancer in Greene's hands, but the numerous reports of Research, 11:811-13, 1951. 5. EICHWALD,E. J.; CHANO,H. Y.; LANDA,M.; EVANS, relative failures with this method (3, 12, 15) dem R. G. Host and Extraneous Factors in Heterologous Tu onstrate that, while this technic may permit mor Transplantation. J. Nat. Cancer Inst., 2:17-81, 1950. initial transplantation of a wide variety of human 6. GREENE,H. S. N. Heterologous Transplantation of a Hu neoplasms, it will not provide investigators with man Fibrosarcoma. Cancer Research, 2:649-54,1942. 7. . The Significance of the HeterotransplanUbility of easily obtained, dependable sources of human Human Cancer. Cancer, 5:1952. tumors in animal hosts. It is pertinent to compare 8. GREENE,H. S. N., and SAXTON,J. A., JR. Uterine Adeno the 3$ years, reported by Dobyns (2), required mata in the Rabbit. I. Clinical History, Pathology, and for eight passages of a thyroid anterior chamber Preliminary Transplantation Experiments. J. Exper. Med., transplant, with the 3$ months required for eight 67:691-707, 1938. 9. KAUFMAN,N.; PRIETO, L. C., JR.; MASON,E. J.; and passages of the Deac-1 tumor reported here. It is KINNET,T. D. The Survival of Human Tumor Tissue on possible that certain tumors might respond well if the Chorio-allantoic Membrane of the Chick Embryo. transplanted initially into the anterior chamber, Am. J. Path., 28:561-62, 1952. with subsequent transfer to the hamster or rat, 10. LEMON,H. M.; Ltrrz, B. R.; POPE, R.; PARSONS,L.; HANDLER,A.H.; and PATT,D. I. Survival and Growth of where technic may be less crucial and growth more Human Tissues Transplanted to Hamster Cheek Pouch. rapid. Science, 115:461-64, 1952. To us, the possibilities of early clinical applica 11. LUTZ,B. R.; FULTON,G. P.; PATT,D. I.; and HANDLER, A. H. The Growth Rate of Tumor Transplants in the tion of this technic are as attractive as the search Cheek Pouch of the Hamster. Cancer Research, 10:231- for other transplantable tumors. Although it may 82, 1950. seem overly optimistic to consider ways and means 12. Srn iLUM:. J. A.; SNELL,A. J.; and FAVATA,B.V. Heterol of destroying the Deac-1 tumor in its animal hosts ogous Ocular Transplantation as a Practical Test for Can at this early date, we are initiating such studies, cer. Cancer, 2:480-90, 1949. 13. TOOLAN,H. W. Successful Subcutaneous Growth and recognizing that the opportunity to test the cor Transplantation of Human Tumors in X-irradiated Labo relation between tumor response in animal and ratory Animals. Proc. Soc. Exper. Biol. & Med., 77:572- human hosts will probably present itself in the 78, 1951. foreseeable future. Should this patient, who is now 14. . The Growth of Human Tumors in Cortisone-treat clinically free of disease, develop a recurrence, ed Laboratory Animals. The Possibility of Obtaining Per manently Transplantable Human Tumors. Cancer Re information on the susceptibility of his cancer to search, 13:389-94, 1953. irradiation, endocrine changes, and antitumor 15. TOWBIN,A. The Heterologous Transplantation of Human drugs would be of immediate interest. Tumors. Cancer Research, 11:716-22, 1051.

W. Bradford Patterson, Rosanna N. Chute and Sheldon C. Sommers

Cancer Res 1954;14:656-659.

Updated version Access the most recent version of this article at: http://cancerres.aacrjournals.org/content/14/9/656

E-mail alerts Sign up to receive free email-alerts related to this article or journal.

Reprints and To order reprints of this article or to subscribe to the journal, contact the AACR Publications Subscriptions Department at [email protected].

Permissions To request permission to re-use all or part of this article, use this link http://cancerres.aacrjournals.org/content/14/9/656. Click on "Request Permissions" which will take you to the Copyright Clearance Center's (CCC) Rightslink site.