Primary culture and hepatocelluar specific protein expression in human amniotic epithelial cells / 上海第二医科大学学报

ABSTRACT

Objective To optimize the method of primary culture for human amniotic epithelial cells (AECs) and detect the expression of hepatocelluar specific proteins in AECs. Methods AECs were obtained by collagenase and trypsin digestion method. 10,20,40 ng/mL epidermal growth factor (EGF) and 10 ng/mL basic fibroblast growth factor were added separately to the culture media,inverted microscope was used to observe the cell growth and proliferation,and the optimal condition for primary culture of AECs was obtained. The primary cells cultured without growth factors were served as controls. The expression of hepatocelluar specific proteins such as albumin,cytokeratin-18 (CK-18),alpha-1 antitrypsin (AAT) and alpha-1 foetoprotein (AFP) in cultured cells and histological sections were detected by immunohistochemical staining. Results The AECs available were relatively pure,with only a few mesenchymal cells. EGF of 10 ng/mL was chosen as an ingredient of culture medium as the growth and proliferation of AECs significantly accelerated with 10 ng/mL EGF. The expression of albumin,CK-18,AAT and AFP was detected in amnion tissues and AECs cultured in vitro. Conclusion Collagenase and trypsin digestion method and culture medium with 10 ng/mL EGF are favourable conditions for primary culture of AECs. Hepatocelluar specific proteins are expressed in human amnion tissues and AECs cultured in vitro,indicating that AECs have some characteristics of hepatocytes.