[CANCER RESEARCH 40. 2500-2506. July 1980] 0008-5472/80/0040-OOOOS02.00 Cell Line Derived from a Metastasis of a Human Testicular Germ Cell Tumor1 David L. Bronson,2 Peter W. Andrews, Davor Solter, Jaroslav Cervenka, Paul H. Lange, and Elwin E. Fraley Departments of Urologie Surgery [D. L. B.. P. H. L., E. E. F.] and Oral Pathology ¡J.C.¡,University of Minnesota College of Health Sciences, Minneapolis. Minnesota 55455. and the Wistar Institute. Philadelphia, Pennsylvania 19104 [P. W. A.. D. S.J ABSTRACT A cell line, designated 833K-E, has been established from a metastasis of a human testicular germ cell tumor that consisted of four histological types of tumor cells. The 833K-E cells have morphological and ultrastructural characteristics of epithelial cells and a hyperdiploid karyotype indicative of their human male origin. The cells grow in agar cultures and produce in nude mice tumors which have the histological features of embryonal carcinoma without differentiated elements. Many of the cells express a stage-specific mouse embryonic antigen, and low levels of the major histocompatibility antigens and ß2- microglobulin also were detected on a large percentage of the cells. A lymphoblastoid cell line (833K-LC) established from the same tumor specimen expresses major histocompatibility antigens and /?2-microglobulin but does not express the embry onic antigen. INTRODUCTION Carcinoma of the testis causes 11 to 13% of the deaths from cancer in North American men between the ages of 15 and 34 years, and more than 90% of the testicular tumors in this age group are classified as germ cell tumors (18). Current evidence suggests that these tumors arise from the primordial germ cells, which line the seminiferous tubules and are the direct precursors of sperm. In the histological classification system proposed by the World Health Organization, testicular germ cell tumors are classified further as EC,3 teratoma, teratocarcinoma (EC with teratoma), choriocarcinoma, yolk sac tumor, and seminoma (20). EC consists of anaplastic, undifferentiated cells that re semble undifferentiated normal cells of an embryo in the earli est stages of development. Approximately 40% of testicular germ cell tumors contain elements of more than one (e.g., teratocarcinoma), or even all, of the histological types of these tumor cells, and it is thought that the other nonseminomatous germ cell tumors are formed by the differentiation of EC cells (14, 18, 20). Thus, malignant transformation of the germ cells leads to the development of seminoma or, by a separate pathway, of EC, and the EC cells differentiate to form chorio carcinoma, yolk sac tumor, or teratoma. In addition, some teratomas contain cartilage, epithelium, and neural tis- ' This investigation was supported in part by Grants CA 15551. CA 10815, and CA 21069 from the National Cancer Institute; T32-GM 07511-03 from the National Institute of General Medical Sciences; HD 12487 from the National Institute of Child Health and Human Development; and 1-301 from the National Foundation-March of Dimes. 2 To whom requests for reprints should be addressed. 3 The abbreviations used are: EC. embryonal carcinoma; HLA, human histo compatibility leukocyte antigens: SSEA-1, stage-specific embryonic antigen; PBS, phosphate-buffered saline (0.9 ITIM CaCI2, 2.7 mM KCI, 1.5 RIM KH2PO4. 138 mM NaCI. and 9.6 mM Na2HPO«.pH 7.4). Received August 21. 1979; accepted April 10. 1980. sue, which suggests the formation by teratoma cells of fully differentiated elements. Much of the information about these tumors was obtained from studies of mouse testicular teratomas, which were first observed by Stevens and Little (28) and were described in detail by Stevens (25) and by Pierce (22). Testicular teratomas occur spontaneously at high frequency in strain 129 mice and can be induced by grafting genital ridges of 12-day embryos or whole embryos into the testes of syngeneic males (26, 27). These teratomas consist of a variety of tissues representing derivatives of all 3 germ layers. Some of the induced and spontaneous mouse teratomas can be serially transplanted in syngeneic adult animals. The trans- plantable tumors also contain EC cells and thus are classified as teratocarcinomas. Several EC cell lines have been estab lished in vitro from these tumors. Some are nullipotential lines, which exhibit little or no tendency to differentiate, whereas others are multipotential lines, which differentiate readily in vitro and in vivo. These EC cells express cell surface (embry onic) antigens that also are detected on undifferentiated cells from other species, including humans, but are not expressed by differentiated cells (reviewed in Rets. 10, 13, and 19). We are aware of reports of only 4 cell lines, Tera-1, Tera-2, SuSa, and NEC-8, that were derived from human testicular germ cell tumors (9, 11, 30). In a previous communication (4), we described ultrastructural observations of the production, although at low frequency, of particles morphologically identi cal to the human placental retrovirus (6, 16, 29) by cells (designated 833K-E cells) established in vitro from an abdom inal metastasis of a human testicular germ cell tumor. This report describes some additional properties of the 833K-E cell line and compares these properties with those of other human and mouse cell lines of similar classification. MATERIALS AND METHODS Clinical History. A right radical orchiectomy was performed in September 1975 on a 19-year-old Caucasian male. Histo- pathological examination of the testis revealed teratoma, EC, seminoma, and foci of choriocarcinoma. Chemotherapy was initiated with methotrexate, cyclophosphamide, and actinomy- cin D, but EC was discovered in a left periaortic lymph node in February 1976, and the patient died 2 months later with wide spread métastases. Tissue Culture. Tissue from an abdominal metastasis, con sisting of choriocarcinoma with elements of EC, teratoma, and seminoma, was obtained at autopsy. The specimen was placed in culture by the coverslip method (8), and cultures were incubated at 35° in Roswell Park Memorial Institute Medium 1640 containing 10% tryptose phosphate broth, 15% heat- inactivated (56°,30 min) fetal bovine serum, 2 mM L-glutamine, 2500 CANCER RESEARCH VOL. 40 Research. on November 25, 2021. © 1980 American Association for Cancer cancerres.aacrjournals.org Downloaded from