[en] Culture conditions for maintaining first trimester human placenta in organ culture, which enhance the secretion of human chorionic gonadotropin (hCG), are described. Nutrient medium, oxygen tension and Gelfoam support matrix influence the synthesis of hCG by these cultures. Placental tissue remained viable for the duration of experiments (12 days) as judged by the incorporation of tritiated thymidine into DNA and the lack of release of incorporated [125I]iododeoxyuridine. Optimal conditions for hCG synthesis in placental organ culture included an atmosphere of 95% air and 5% Co2 (approximately 20% O2), CMRL 1066 medium containing fetal human or bovine serum, insulin, hydrocortisone and retinal acetate. Multiple pieces of placenta could be cultured in the same dish with an additive effect on hCG secretion. The functional responsiveness of these placental cultures was demonstrated by modulation of hCG synthesis with theophylline and 3'5' dibutyryl cyclic AMP.
Disciplines :
Reproductive medicine (gynecology, andrology, obstetrics)
Author, co-author :
Huot, R. I.
Foidart, Jean-Michel ; Université de Liège - ULiège > Département des sciences cliniques > Gynécologie - Obstétrique - Labo de biologie des tumeurs et du développement
Stromberg, K.
Language :
English
Title :
Effects of culture conditions on the synthesis of human chorionic gonadotropin by placental organ cultures.
Publication date :
1979
Journal title :
In Vitro
ISSN :
0073-5655
Publisher :
Tissue Culture Association, Rockville, United States - Maryland
Thiede H.A., Rudolph J.H. (1961) A method for obtaining monolayer cultures of human fetal cells from term placentas. Proc. Soc. Exp. Med. Biol. 107:565-569.
Taylor P.V., Hancock K.W. (1973) Viability of human trophoblast in vitro. J. Obstet. Gynaecol. Br. Commonw. 80:834-838.
Hall C.G., James T.E., Goodyer C., Branchaud C., Gudya H., Giroud C.J.P. (1977) Short term tissue culture of human midterm and term placenta. Parameters of hormonogenesis. Steroids 30:569-580.
Stromberg K., Azizkhan J.C., Speeg K.V. (1978) Isolation of functional human trophoblast cells and partial characterization in primary cell culture. In Vitro 13:631-638.
Chung H.K., McLimans W.F., Horoszewicz J., Hreshchyshyn M.M. (1969) In vitro studies of human trophoblast. Am. J. Obstet. Gynecol. 104:945-952.
Stoner G.D., Harris C.C., Autrup H., Trump B.F., Kingsbury E.W., Meyers G.A. (1978) Explant culture of human peripheral lung. I. Metabolism of benzo(a)pyrene. Lab. Invest. 38:685-692.
Speeg K.V., Azizkhan J.C., Stromberg K. (1976) The stimulation by methotrexate of human chorionic gonadotropin and placental alkaline phosphatase in cultured choriocarcinoma cells. Cancer Res. 36:4570-4576.
Lowry O.H., Rosenbrough N.J., Farr A.C., Randall R.J. (1951) Protein measurement with Folin phenol reagent. J. Biol. Chem. 193:265-275.
Fidler I.J. (1970) Metastasis: Quantitative analysis of distribution and fate of tumor emboli labeled with125I-5-iodo-2′-deoxyuridine. J. Nat. Cancer Inst. 45:773-782.
Liotta L.A., Vembu D., Saini R.K., Boone C. (1978) In vivo monitoring of the death rate of artificial murine pulmonary metastases. Cancer Res. 38:1231-1236.
Hofer K.G., Hofer M. (1971) Kinetics of proliferation, migration and death of L1210 ascites cells. Cancer Res. 31:402-408.
Hughes W.L., Commerford S.L., Gitlin D. (1964) Deoxyribonucleic acid metabolism in vivo: Cell proliferation and death as measured by incorporation and elimination of iododeoxyuridine. Fed. Proc. 23:640-649.
Commerford S.L. (1965) Biological stability of 5-iodo-2-deoxyuridine labeled with iodine 125 after its incorporation into deoxyribonucleic acid of the mouse. Nature 206:949-950.
Feinendegen L.E., Bond V.P., Hughes W.L. (1966) 125IUDR (5-iodo-2-deoxyuridine) in autoradiographic studies of cell proliferation. Exp. Cell Res. 43:107-119.
Hussa R.O., Story M.T., Pattillo R.A. (1974) Cyclic adenosine monophosphate stimulates secretion of human chorionic gonadotropin and estrogens by human trophoblast in vitro. J. Clin. Endocrinol. Metab. 38:338-340.
Handwerger S., Barrett J., Tyrey L., Schomberg D. (1974) Differential effects of cyclic adenosine monophosphate on the secretion of human placental lactogen and human chorionic gonadotropin. J. Clin. Endocrinol. Metab. 36:1268-1270.
Hussa R.O., Story M.T., Pattillo R.A., Kemp R.G. (1977) Effect of cyclic 3′∶5′-AMP derivatives, prostaglandins, and related agents on human chorionic gonadotropin secretion in human malignant trophoblast in culture. In Vitro 13:443-449.
Azizkhan J.C., Speeg K.V., Stromberg K., Goode D.M. (1979) Stimulation of human chorionic gonadotropin synthesis in the JAR line of choriocarcinoma cells by inhibitors of DNA synthesis. Cancer Res. 39:1952-1959.
Gough E.D., Lowe M.C., Juchau M.R. (1975) Human placental aryl hydrocarbon hydroxylase: Studies with fluorescence histochemistry. J. Nat. Cancer Inst. 54:819-824.
