人胎盘的生殖内分泌学研究——Ⅱ.细胞滋养层细胞的激素分泌功能

本文采用了无血清培养技术,研究了细胞滋养层细胞在体外分泌hCG、孕酮、雌激素、cGnRH和β-内啡肽的功能,在8天的培养中发现,细胞滋养层细胞在培养24h去除内源激素后能持续地分泌少量的hCG;与此不同,孕酮的分泌量逐日上升,第6天的孕酮分泌量为第1天的两倍;在外加雌激素的前身物雄烯二酮的情况下,细胞能合成雌激素,其产量亦是逐日上升,反映出细胞芳香化酶活性的不断提高,这种酶活性的提高与雄烯二酮的存在与否无关;在两种滋养层细胞培养中,唯有细胞滋养层细胞培养液中测得有cGnRH,产量以第2天为最高,接近4pg/10~5个细胞,合体滋养层细胞在离体下能分泌β-内啡肽,但其产量逐日下降,与此相反,细胞滋养层细胞的培养液中不但测到β-内啡肽的存在而且其含量在培养4—6天间明显上升。     

STUDY ON REPRODUCTIVE ENDOCRINOLOGY OF HUMAN PLACENTA (Ⅱ)——HORMONE SECRETING ACTIVITY OF CYTO-TROPHOBLAST CELLS

The capability of cytotrophoblast cells to produce hCG, progesterone, estrogen, cGnRH and β-endorphin in vitro has been demonstrated in serum-free culture medium. Before experiment, a 24-h preculture was carried out in order to remove the endogenous hormones of the tissue. During a period of 8 days' culture, the cytotrophoblast cells could constantly produce a small amount of hCG. The production of progesterone rose rapidly and became doubled within six days. The estrogen secretion showed a similar pattern in the presence of androstenedione, a precursor of estrogen, indicating the elevation of aromatase activity in the cells. The elevation of the enzyme activity has been further demonstrated not to be induced by androstenedione. In both cytotrophoblast and syncytiotrophoblast cell cultures, cGnRH was only detected in the culture of cytotrophoblast cells, with a value up to 4 pg/105 cells/24 h. However, β-endorphin was identified both in cytotrophoblast and syncytiotrophoblast cells. Its content increa     

STUDY ON REPRODUCTIVE ENDOCRINOLOGY OF HUMAN PLACENTA--CULTURE OF HIGHLY PURIFIED CYTOTROPHOBLAST CELL IN SERUM-FREE HORMONE SUPPLEMENTED MEDIUM

A new method of long-term culture of cytotrophoblast cells in serum-free medium has been developed. Cytotrophoblast cells were isolated with cold trypsin and purified by unit gravity sedimentation through BSA density gradients. The cells were cultured in the FD medium with supplement of EGF, insulin, transferrin and sodium selenite. They could survive over three weeks. The results showed that both EGF and insulin stimulated hCG and progesterone secretion and that sodium selenite elevated hCG output but not progesterone secretion. Transferrin produced synergistic effect with EGF and insulin on hCG and progesterone secretion but it was ineffective when used alone. This study demonstrates that the four growth factors mentioned above are essential for the survival of cytotrophoblast cells in vitro. It is therefore suggested that EGF, insulin and selenium may possibly be involved in the regulation of hCG and progesterone secretion in the human placenta.     

人胎盘的生殖内分泌学研究——Ⅰ.人细胞滋养层细胞的无血清培养与生长因子的作用

为了研究胎盘的激素分泌及其调节功能,作者发展了一种简便分离细胞滋养层细胞和用无血清培养的实验模型,研究了上皮生长因子、胰岛素、硒及转铁蛋白等生长因子对细胞滋养层细胞的生存和激素分泌的影响。结果显示在含生长因子的培养液中细胞可生存3周;上皮生长因子和胰岛素对hCG和孕酮的分泌都有刺激作用,硒只影响hCG的分泌,转铁蛋白的作用不明显,但是转铁蛋白与另外两种因子合用时呈现协同作用。本研究说明上述的4种因子和胶原是维持细胞滋养层细胞生存不可缺少的因素,同时也发现上皮生长因子、胰岛素和硒还参与了正常人胎盘激素分泌的调控。