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Induction of the differentiation of porcine bone marrow mesenchymal stem cells into premature hepatocyte-like cells in an indirect coculture system with primary hepatocytes

Animal Cells and Systems 2020년 24권 5호 p.289 ~ 298
Ullah Imran, 서강민, 위하연, 김영임, 이승훈, 옥선아,
소속 상세정보
 ( Ullah Imran ) - Rural Development Administration National Institute of Animal Science Animal Biotechnology Division
서강민 ( Seo Kang-Min ) - Rural Development Administration National Institute of Animal Science Animal Biotechnology Division
위하연 ( Wi Ha-Yeon ) - Rural Development Administration National Institute of Animal Science Animal Biotechnology Division
김영임 ( Kim Young-Im ) - Rural Development Administration National Institute of Animal Science Animal Biotechnology Division
이승훈 ( Lee Seung-Hoon ) - Rural Development Administration National Institute of Animal Science Animal Biotechnology Division
옥선아 ( Ock Sun-A ) - Rural Development Administration National Institute of Animal Science Animal Biotechnology Division

Abstract


Liver transplantation is currently the only option for patients with end-stage liver disease. Thus, other alternate therapeutic strategies are needed. Bone marrow mesenchymal stem cells (BM-MSCs) are nonhematopoietic cells present in the bone marrow stroma that serve as precursors cells for various other cells. In this study, we evaluated the differentiation of porcine BM-MSCs into hepatocyte-like cells using three types of culture systems: hepatic induction medium (HIM), HIM/primary hepatocyte culture supernatant (HCS; 1:1 ratio), and a hepatocyte coculture system (HCCS; primary hepatocytes in the upper chamber, and BM-MSCs in the lower chamber). Primary hepatocytes were isolated from anesthetized healthy 1-month-old pigs by enzymatic digestion. Hepatic-specific marker expression (albumin [ALB], transferrin [TF], α-fetoprotein [AFP]), glycogen storage, low-density lipoprotein, and indocyanine green uptake were evaluated. Upregulation of hepatic-specific markers (ALB, TF, and AFP) was observed by real-time polymerase chain reaction in the HCCS group. Periodic acid-Schiff staining revealed enhanced glycogen storage in hepatocyte-like cells from the HCCS group compared with that from the HIM/HCS group. Furthermore, hepatocyte like-cells in the HCCS group showed improved LDL and ICG uptake than those in the other groups. Overall, our current study revealed that indirect coculture of primary hepatocytes and BM-MSCs enhanced the differentiation efficacy of BM-MSCs into hepatocyte-like cells by unknown useful soluble factors, including paracrine factors.

키워드

Mesenchymal stem cells; bone marrow mesenchymal stem cells; hepatocyte-like cells; coculture; Transwell system

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