Several Important in Vitro Improvements in the Amplification, Differentiation and Tracing of Fetal Liver Stem/progenitor Cells

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2012 Oct 12

PMID 23056632

Citations 1

Authors

Wei-hui Liu

Zheng-Cai Liu

Nan You

Ning Zhang

Tao Wang

Zhen-Bin Gong

Hong-Bao Liu

Ke-Feng Dou

Affiliations

Soon will be listed here.

Abstract

Objective: We previously isolated fetal liver stem/progenitor cells (FLSPCs), but there is an urgent need to properly amplify FLSPCs, effectively induce FLSPCs differentiation, and steadily trace FLSPCs for in vivo therapeutic investigation.

Methods: FLSPCs were maintained in vitro as adherent culture or soft agar culture for large-scale amplification. To direct the differentiation of FLSPCs into hepatocytes, FLSPCs were randomly divided into four groups: control, 1% DMSO-treated, 20 ng/ml HGF-treated and 1% DMSO+20 ng/ml HGF-treated. To trace FLSPCs, the GFP gene was introduced into FLSPCs by liposome-mediated transfection.

Results: For amplifying FLSPCs, the soft agar culture were more suitable than the adherent culture, because the soft agar culture obtained more homogeneous cells. These cells were with high nuclear:cytoplasmic ratio, few cell organelles, high expression of CD90.1 and CD49f, and strong alkaline phosphatase staining. For inducing FLSPCs differentiation, treatment with HGF+DMSO was most effective (P<0.05), which was strongly supported by the typical morphological change and the significant decrease of OV-6 positive cells (P<0.01). In addition, the time of indocyanine green elimination, the percentage of glycogen synthetic cells, and the expressions of ALB, G-6-P, CK-8, CK-18 and CYP450-3A1 in HGF+DMSO-treated group were higher than in any other group. For tracing FLSPCs, after the selection of stable FLSPC transfectants, GFP expression continued over successive generations.

Conclusions: FLSPCs can properly self-renew in soft agar culture and effectively differentiate into hepatocyte-like cells by HGF+DMSO induction, and they can be reliably traced by GFP expression.

Citing Articles

Unbalanced distribution of materials: the art of giving rise to hepatocytes from liver stem/progenitor cells.

Liu W, Ren L, Chen T, You N, Liu L, Wang T J Cell Mol Med. 2013; 18(1):1-14.

PMID: 24286303 PMC: 3916112. DOI: 10.1111/jcmm.12183.

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