Umbilical cord mesenchymal stem cells support the in vitro growth of primary mouse hepatocytes by a paracrine mechanism

doi:10.3969/j.issn.2095-4344.1785

Abstract

Abstract:

BACKGROUND: Hepatocyte transplantation is a very effective alternative treatment for patients with advanced liver failure. It is of very important clinical significance to expand and culture hepatocytes in vitro.
OBJECTIVE: To explore the regulatory effect of human umbilical cord mesenchymal stem cells on the function of primary mouse hepatocytes under in vitro conditions.
METHODS: Umbilical cord mesenchymal stem cells were co-cultured with primary mouse hepatocytes in a non-contacted way (co-culture group). Umbilical cord mesenchymal stem cell supernatant was collected and co-cultured with primary mouse hepatocytes (conditioned medium). Primary mouse hepatocytes were cultured in conventional culture medium containiing low-dose DMEM+10% fetal bovine serum as control group. Cell counting kit-8 method was used to detect the proliferation of three groups of hepatocytes. RT-PCR was used to detect the expression of hepatocyte-related genes. Indocyanine green detection was used to detect the function of hepatocytes. ELISA method was used to detect the secretion of albumin and urea. The implementation of the study was in line with the relevant ethical requirements of Kashgar Region First Hospital (ethical approval No. 01).
RESULTS AND CONCLUSION: Compared with the control group, umbilical cord mesenchymal stem cells could significantly retain the uptake ability of mouse hepatocytes (96 hours of culture) and promote their proliferation (48, 72, and 96 hours of culture) by non-contact co-culture (P < 0.05) or supernatant culture (P < 0.05). Compared with the control group, the primary mouse hepatocytes in the co-culture group and the conditioned medium group had significantly stronger albumin and urea secretion ability (P < 0.05). RT-PCR detection of related genes revealed higher expression of hepatocyte growth factor, basic fibroblast growth factor and album mRNAs in the co-culture group and conditioned medium group than the control group (P < 0.05). However, we observed similar results in the expression of the cytochrome gene Cyp2b9 in the three groups (P > 0.05). Overall, umbilical cord mesenchymal stem cells can preserve the function of primary mouse hepatocytes in vitro by the paracrine mechanism and promote their proliferation. It is a new application strategy for artificially expanding hepatocytes in vitro.

Key words: human umbilical cord mesenchymal stem cells, hepatocytes, co-culture, paracrine, hepatocyte growth factor, liver function, Natural Science Foundation of Xinjiang Uygur Autonomous Region

CLC Number:

Wu Yuanquan, Wu Jianhua, Wang Liang, Lin Yang. Umbilical cord mesenchymal stem cells support the in vitro growth of primary mouse hepatocytes by a paracrine mechanism[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(25): 4031-4036.

Figures/Tables 3

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