THE AVAILABILITY OF YOUNG CHICKS FOR TUMOR TRANSPLANTATION

Transcription

1 THE AVAILABILITY OF YOUNG CHICKS FOR TUMOR TRANSPLANTATION KANEMATSU SUGIURA (From thp Huntington Fund for Cancer Research, Memorial Hospital, and the Harrirnan Research Laboratory, the Roosevelt Hospital, New York) The discovery of a transplantable chicken sarcoma in 1910 by Rous (1) and Fuzinami and Inamoto () has resulted in the use of large numbers of chickens for experimental and routine tumor transplantation. A recent report of Gye (3) upon the etiology of cancer, has revived interest in the sarcoma of fowls and has stimulated the use of chickens in several fields of cancer research (-10). It is generally agreed that tumor susceptibility varies among hens. The variable rate of growt3h of inoculated tumor tissue in hens and the occasional failure of these tumors to proliferate when transplanted, has been an annoying hindrance to progress in this type of tumor research. A suitable and reliable soil for tumor growth is desirable. Transplantation experiments with the mammalian tumors indicate that young animals are more suitable for successful inoculation than old animals. On account of this, it was thought of value to investigate the degree of adaptability of young chicks in tumor transplantation. Should the tumor show a dependable growth in young chicks the expense of investigation with chicken sarcoma might be greatly reduced. EXPERIMENTAL P.RT In the present study we used Barred Plymouth Rock chicks, whose ages varied from one to thirty days at the time of the primary tumor transplantation. The chicks, generally 1 in number were kept in wire cages, 1 X 1 x 1 inches. The floor of each cage was covered with sawdust. The room in which these cages were kept was well ventilated, lighted and maintained at a moderate and equable 8 1

2 8 KANEMATSU SUGIURA temperature. The initial diet consisted of Conkey's starting feed with occasional feedings of sweet milk. Later the chicks were given lettuce and grains supplemented by broken charcoal, and granite. They had constant access to fresh tap water. A number of growth curves of normal chicks andl of tumorbearing chicks are shown in Fig. 1. FIG. 1. COMPARIRON OF (:ROWTI< IN BODY WICIGHT OF FORMAL (1) ANI) OF TUMOR-BEARINU (, 3 AND ) HARRED PLYMOPTH ROCK CHICKS FROM A DAY AFTER HATCHING. At the poiiit marked by the :mr(iw ( ) the experimental chicks were inocriltited with Roiis chicken sarcoma. The crosses indicate the time of death. It will be noted that with the continued growth of the tumor, the body weight of an inoculated chick increases slowly until its death, but its rate of growth is not so rapid as that of the normal chick. Throughout the experiments we used fresh growth of Rous chicken sarcoma No. 1. The growth of the first generation was obtained from a desiccated and pulverized tumor tissue which was kindly furnished by Dr. I'eyton Rous of the Rockefeller Institute for Medical Research. Tumor tissue weighing about milligrams was selected from a slightly hemorrhagic area of a tumor two weeks old, and was inoculated into the pectoral muscles of the chicks by a small trocar, the diameter of which was approximately 1. mm. Observations upon the normal implantations are presented in

3 YOUNG CHICKS FOR TUMOR TRANSPLANTATION 83 Table I. The data are arranged according to the ages of the chicks into which the pieces of tumor tissue were inoculated. - Experiment No TABLE I Results oj Transplanting Rous Chicken Sarcoma in Young Chicks (Barred Plymouth Rocks) at Iliflerent Ages Number of Chick8 Urrd Ages of Chicks in Days wbcr Tumor Tissue was Inoculatec I Tumor Tissue Used c - RCH 7c - RCH 1) - R C8 13 H B - 13 CS 11 XA RCS ' - R CS 3D 31) B R CS BC Percentage of Tumor Takes 0 7 We can see, from the results in Table I, that the age of the chicks does not affect the percentage of tumor takes, since in most experiments, perfect transplantation resulted. In spite of the small size and undeveloped pectoral muscles of these chicks the transplants grew rapidly, metastasized readily and killed the chicks quickly. Once this tumor was established

4 8 KANEMATSU SUGIURA and growing in chicks it never underwent spontaneous retrogression. Many of the chicks which were inoculated with the tumor tissue at an age of less than 8 days, died in about 1 days after this implantation. As these chicks grew ollder their powers of resistance increased. The sarcoma did not kill the 1 and 30 day old chicks before and 30 days respectively after tumor implantation. In this respect the behavior of older chicks is similar to that of young adult hens or pullets. Forty-two per cent of the autopsied chicks dying from the neoplastic disease, showed metastases in the lungs, :and 38 per cent showed petechial hemorrhages in the liver. Aside from these two immediate causes of death, the tumor affects the nutritional state of its host and apparently finally kills it by nutritional failure. In diminishing order of frequency, tumor metastases occurred in the lungs, liver, heart, kidneys, and spleen. Both male and female chicks were found equally susceptible to inoculation with the Rous chicken sarcoma. Among 71 chicks used in the normal transplantation experiments shown in Table I, 0 were males and 31 were females. The sarcoma grows relatively faster and metastasizes quicker in healthy growing chicks than in young adult, hens. In consequence, the tumor reaches a greater size in the hen than in the chick. It is of interest to compare the beh:tvior of the chicken sarcoma with that of malignant mammalian tumors; e.g., the Flexner-Jobling rat carcinoma and the Bashford mouse carcinoma 3 in suckling rats and mice respectively, grew progressively and almost equally in size with those in adult animals (11). The re-implantation with similar tumor tissue within a short time after the primary implantation did not affect the takes of both primary and secondary implants. This result differs somewhat from that obtained with mammalian neoplasms (1) indicating that different tumor parenchymata vary in their power of inducing resistance. Another point to be noted in this connection is that the

5 I ~ ~ ~ YOUNG CHICKS FOR TUMOR TRANSPLANTATION 8 tumor takes and their subsequent rate of growth in hens were irregular. There were instances where tumor development in hens required a period of three weeks in order to reach a measurable size. Young chicks tolerated quite well the mechanical injuries which are involved in the tumor inoculation, and very seldom died as a result of the trauma. On account of their small size and high susceptibility to Rous chicken sarcoma they may be used to advantage in many experiments. TRANSPLANTATION OF ROUS CHICKEN SARCOMA IN CHICKS OF DIFFERENT BREEDS In connection with the present study we determined the degree of inoculability of Rous chicken sarcoma in breeds of chicks other than the Plymouth Rock in which the original tumor occurred. TABLE I1 Results of Transplanting Rous Chickan Sarcoma in Young Chicks of Digrrent Breeds Experiment No. l ,._ , Kind of Rrrcds White JTyandottes White Wy an do t tes IJlack Jersey Giants Black.Jersey Giants White Leghorns White Leghorns Rhode Island Reds Rhode Island Reds Rhode Island Reds Number If Chicks Used 3 Agrs of Chirk, in h.vs when Tumoi Tissue \\as Inoculated 1s 1 Tumor Tissue LTscd - Itcs KCS 9n XA 9B RCS 9U 10A 'errentage if Tumor Takes I00

6 8 KANEMATSU SUGIURA The tumor tissues used in these experiments were selected from tumors which have been passed through several Barred Plymouth Rock chicks. The data obtained from these experiments are given in Table 11. The above results clearly show that the Rous chicken sarcoma grows well in the different breeds of fowls commonly seen on American farms. The frequency and the site of metastatic deposits in these chicks were similar to those found in the Barred Plymouth Rock chicks. However, the rate of tumor growth in White Wyandottes was somewhat slower than was found in other breeds. SUMMARY The susceptibility of young chicks (one to 30 da,ys old) to Rous chicken sarcoma No. 1 was studied under laboratory conditions. It was found that the Barred Plymouth Rocks, White Wyandottes, Black Jersey Giants, White Leghorns, and Rhode Island Reds were equally susceptible to the tumor growth. Re-inoculation within short periods did not affect the normal growth of the first or second implants. The small size of young chicks, the ease in caring for large numbers of them, the low cost and upkeep as compared with hens, and their high and uniform susceptibility to Rous chicken sarcoma are factors to be considered in using them for experimental tumor studies. REFEltENCES 1. lloua, P.: J. Exp. Med., 1910, xii, 9.. FUZINAMI, A,, AND INAMOTO, K.: Gann, 1911, v, GYE, W. E.: IAuncet, 19, ccix, MURPHY, J. U.: J. A. M. A., 19, Ixxxvi, MUELLBR, J. H.: Proc. Soc. Exp. I3iol. and Med., 19, xxiii, 70.. FLU, P. C.: C. It. Sac. Bid., Paris, 19, xcix, HARDE, E.: J. Tropical Med. and Hygiene, 193, xxix, HARKINS, M. J., SCHAMBIGRG, J. F., AND KOLMER, J. A,: J. Cancer Res., 19, x,. 9. CUTLER, M.: J. Cancer Res., 19, x, NAKAHARA, w.: Gunn, 19, XX, SUGIURA, I<.: J. Cancer Res., 19, viii, SUGIURA, I<., ANU UENEDICT, 8. R.: J. Cancer Res., 19, x, 1%