Species - Tissue - Features - Reference - Growth - Passage - Medium - Freeze - ATCC

Species: human

Tissue: Choriocarcinoma

Features: Gonadotropin, lactogen and steroid secreting

Reference: see ATCC

Mode of Growth:

Method of Passage:

Culture Medium: 12K modification of Ham's medium, 85%; FBS, 15%

Freeze Medium: Culture medium, 95%; dimethyl sulfoxide (DMSO), 5%

ATCC: ATCC CCL-98, BeWo (Choriocarcinoma, human) Gonadotropin, lactogen and steroid secreting. Current medium for propagation: F12K modification of Ham's medium, 85%; FBS, 15%. The BeWo trophoblastic cell line, the first human endocrine cell type to be maintained in continuous cultivation, was initiated in June, 1966 by R.A. Pattillo and G.O. Gey from a malignant gestational choriocarcinoma of the fetal placenta (Cancer Res. 28: 1231, 1968; Science (Washington, DC) 159: 1467, 1968). The tumor was originally transplanted to the hamster cheek pouch by R. Hertz where it was maintained for 304 serial transfers over a period of 8 years before establishment in vitro (Proc. Soc. Exp. Biol. Med. 102: 77, 1959). The original BeWo culture was established from a parabiotic combination of the choriocarcinoma and human decidual tissue embedded on a collagen matrix. The initial medium consisted of Waymouth's MB 752/1 diluted with Gey's BSS, 70%; placental-cord serum, 30%. Homogeneous populations of trophoblast cells were developed after a period of several months by means of microsurgically excising and discarding the fibroblastic and other non-tumorous areas. Gradual adaptation to growth on glass and plastic flasks in Waymouth's MB 752/1, 50%; Gey's BSS, 40%; newborn calf serum, 10% was accomplished over a 5-month period. The BeWo cell line has been shown to secrete the spectrum of placental hormones including human chorionic gonadotropin (hCG), polypeptide hormones, human placental lactogen (hPL), estrogenic and progestational steroids, estrone, estradiol, estriol, and progesterone (In Vitro (Rockville) 6: 205, 1970). The cells synthesize hCG up to a rate of 1000 IU/10(8) cells/24 hours (Ann.N.Y. Acad. Sci. 172: 288, 1971). Cellular differentiation of this stem cell type occurs when hormonal precursors are added to the incubation medium: 35-40% conversion of estrogenic precursors to estrone and estradiol was observed (J. Clin. Endocrinol. Metab. 34: 59-61, 1972). The predominant glycolytic metabolism exhibited by these cells and a high carbon dioxide production and rapid drop in pH, necessitates daily fluid renewals or addition of sufficient culture medium to last 36-48 hours to prevent glucose exhaustion. In culture, approximately 10 million cells subcultured one to three at weekly intervals utilize 1 mg of glucose per hour from the culture medium. Variations in the level of glucose contained in the culture medium exert profound changes on cell growth and glycogen content. A glucose concentration of approximately 300-340 mg percent has been found to be optimal for the cells. The cell line was submitted to the American Type Culture Collection in the 187th passage. DESCRIPTION OF REPOSITORY REFERENCE SEED STOCK: Number of Serial Subcultures from Tissue of Origin: 191. Freeze Medium: Culture medium, 95%; dimethyl sulfoxide (DMSO), 5%;
antibiotic-free. Viability: Approximately 80% (dye exclusion). Culture Medium: Waymouth's MB 752/1, 50%; Gey's BSS, 40%; newborn calf serum, 10%; antibiotic-free. Growth Characteristics of Thawed Cells: An inoculum of 6-7 X 10(5) viable cells in 4 ml of the above culture medium per 25 cm2 flask yields a 3- to 5-fold increase within 7 days at 37C, provided the medium is renewed daily and the total volume of medium per flask is increased by 1 ml each day. Plating Efficiency: Not measurable in the above culture medium. Morphology: Epithelial-like. Karyology: Chromosome Frequency Distribution 75 Cells: 2n = 46 Cells: 1 1 2 3 3 4 3 8 5 17 1 3 4 5 4 1 10 Chromosomes: 71-73-76-79-81 82 83 84 85 86 87 88 89 90-92 93 97-178 Stemline number is hypotetraploid. Karyotype relatively stable within stemline number. Sterility: Tests for mycoplasma, bacteria, and fungi were negative. Species: Confirmed as human by the cytotoxic-antibody dye-exclusion test. Virus Susceptibility: Susceptible to poliovirus type 3, and vesicular stomatitis (Indiana Strain) virus. Reverse Transcriptase: Not detected. Hormone Production: At passage 195, an average of 64 + or - 3 (S.E.) IU of hCG were secreted per 10(6) cells per 24 hours. At passage 198, an average of 35 + or - 3 (S.E.) nanograms of progesterone were secreted per mg of cell protein per hour. Both hCG and progesterone were measured by radioimmunoassay. Isoenzymes: G6PD type B. Submitted by: R.A. Pattillo, Department of Gynecology and Obstetrics, The Medical College of Wisconsin, Milwaukee, WI. Prepared and characterized by: ATCC, Rockville, MD. Price Code: J 

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