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用胶原酶直接消化人胚肝组织小块,可以简便、快速地获得大量活率较高和较纯的肝上皮样细胞。在低钙、低血清浓度加有多种因素的培养液中,胚肝上皮样细胞可以体外培养一月以上。原代培养的胚肝上皮样细胞一次传代后,仍可保持一定的生长能力。表皮生长因子、霍乱毒素、转铁蛋白及肝细胞生长因子等均对培养的胚肝上皮样细胞具有一定的生长刺激作用。本文还用r-GT染色、AFP和白蛋白测定,以及~3H-TdR掺入等指标观察了培养过程中胚肝上皮样细胞的生长、增殖和生物学功能。人胚肝上皮样细胞的较长期体外培养,可望成为研究肝细胞生长、分化及癌变的一个有用的实验模型。
Direct digestion of human embryonic liver tissue with collagenase small pieces, you can easily and quickly access to a large number of high survival and more pure liver epithelial-like cells. In low calcium, low serum concentration plus a variety of factors in the culture medium, embryo liver epithelial-like cells can be cultured in vitro for more than a month. Primary culture of embryonic liver epithelial-like cells after a passage, can still maintain a certain ability to grow. Epidermal growth factor, cholera toxin, transferrin and hepatocyte growth factor all have certain growth stimulating effects on cultured embryonic liver epithelial-like cells. In this paper, we also observed the growth, proliferation and biological function of embryonic liver epithelial-like cells during culture by r-GT staining, AFP and albumin assay, and 3H-TdR incorporation. Prolonged in vitro culture of human embryonic liver epithelial cells is expected to become a useful experimental model for the study of hepatocyte growth, differentiation and carcinogenesis.