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Inducing differentiation of human urine‐derived stem cells into hepatocyte‐like cells by coculturing with human hepatocyte L02 cells
Author(s) -
Zhou Ming,
Shen Liangliang,
Qiao Yinggu,
Sun Zhenxiao
Publication year - 2020
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.29301
Subject(s) - hepatocyte , cd90 , microbiology and biotechnology , flow cytometry , stem cell , cellular differentiation , biology , cd34 , chemistry , cd44 , cell , in vitro , biochemistry , gene
Objectives To investigate the possibility of inducing differentiation of human urine‐derived stem cells (hUSCs) into hepatocyte‐like cells by coculturing with human hepatocyte L02 cells in vitro. Methods HUSCs were isolated from fresh urine samples collected from healthy adult volunteers by centrifugation. Cells were observed under an inverted phase contrast microscope, and proliferative activity was determined by 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide assay. Stem cell surface markers were detected by flow cytometry. HUSCs were induced to differentiate into hepatocyte‐like cells by coculturing with human hepatocyte L02 cells, which were confirmed by cellular morphology, messenger RNA expression of albumin (ALB), α‐fetoprotein (AFP) and hepatocyte cytochrome P450 (CYP450) analyzed with quantitative reverse transcription polymerase chain reaction and the expression of glycogen detected by glycogen staining kits at 5, 10, and 15 days after coculturing. Results HUSCs from urine were successfully isolated and cultured in vitro. At passages 3, the growth curve of hUSCs was S‐shaped with good proliferation activity. Mesenchymal stem cell surface markers CD44 and CD90 were detected positive by flow cytometry. CD31 for endothelial cells and CD34 for hematopoietic stem cell markers were not detected. HUSCs gained the cellular morphology and function of hepatocyte cells including higher expression of several hepatocyte‐specific genes such as ALB and some CYP450, lower expression of AFP and positive glycogen expression ( P < .05) in coculturing with human hepatocyte L02 cells for 10‐15d. Conclusions HUSCs can be induced to differentiate into hepatocyte‐like cells by coculturing with human hepatocyte L02 cells for a certain number of days.