Long-term culture of rat hepatocytes using human amniotic membrane as a culture substrate

Kyoichiro Maekawa; Koji Natsuda; Masaaki Hidaka; Masafumi Uematsu; Akihiko Soyama; Takanobu Hara; Mitsuhisa Takatsuki; Kazuhiro Nagai; Kiyonori Miura; Susumu Eguchi

doi:10.1016/j.reth.2021.09.002

Long-term culture of rat hepatocytes using human amniotic membrane as a culture substrate

Regen Ther. 2021 Sep 28:18:384-390. doi: 10.1016/j.reth.2021.09.002. eCollection 2021 Dec.

Authors

Affiliations

¹ Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
² Department of Ophthalmology and Visual Sciences, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
³ Department of Digestive and General Surgery, Graduate School of Medicine, University of the Ryukyus, Okinawa, Japan.
⁴ Transfusion and Cell Therapy Unit, Nagasaki University Hospital, Nagasaki, Japan.
⁵ Department of Obstetrics and Gynecology, Nagasaki University Graduate School of BioMedical Sciences, Nagasaki, Japan.

Abstract

Amniotic membrane is attracting attention as a new material for regenerative medicine. We herein report that the culture of primary rat hepatocytes on human amniotic membrane maintained their morphology and their production of albumin for at least two months. Human amniotic membrane was collected during planned cesarean section and kept frozen until usage. Primary rat hepatocytes were plated on human amniotic membrane. Hepatocytes accumulated as colonies on amniotic membrane, and their rat albumin level was maintained for two months. Their three-dimensional structure on extracellular matrix, which is abundant in amniotic membranes might influence the maintenance of the hepatocyte-specific function.

Keywords: AM, amniotic membrane; Albumin synthesis; DMSO, dimethyl sulfoxide; EGF, epidermal growth factor; ELISA, enzyme-linked immunosorbent assay; FBS, fetal bovine serum; HBV, hepatitis-B virus; HCV, hepatitis-C virus; HGF, hepatocyte growth factor; HIV, human immunodeficiency virus; HTLV-1, human T-cell leukemia virus type 1; Human amniotic membrane; LT, liver transplantation; PBS, phosphate-buffered saline; Rat hepatocyte.