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Stem Cells Dev. 2017 Aug 1;26(15):1111-1120. doi: 10.1089/scd.2017.0026. Epub 2017 Apr 25.

Generation of Canine Induced Extraembryonic Endoderm-Like Cell Line That Forms Both Extraembryonic and Embryonic Endoderm Derivatives.

Author information

1
1 Department of Advanced Pathobiology, Graduate School of Life and Environmental Sciences, Osaka Prefecture University , Osaka, Japan .
2
2 Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University , Kyoto, Japan .

Abstract

Extraembryonic endoderm (XEN) cells are stem cell lines derived from primitive endoderm cells of inner cell mass in blastocysts. These cells have self-renewal properties and differentiate into visceral endoderm (VE) and parietal endoderm (PE) of the yolk sac. Recently, it has been reported that XEN cells can contribute to fetal embryonic endoderm, and their unique potency has been evaluated. In this study, we have described the induction and characterization of new canine stem cell lines that closely resemble to XEN cells. These cells, which we designated canine induced XEN (ciXEN)-like cells, were induced from canine embryonic fibroblasts by introducing four transgenes. ciXEN-like cells expressed XEN markers, which could be maintained over 50 passages in N2B27 medium supplemented with inhibitors of mitogen-activated protein kinase p38 and transforming growth factor-beta 1. Our ciXEN-like cells were maintained without transgene expression and exhibited upregulated expression of VE and PE markers in feeder-free conditions. The cells differentiated from ciXEN-like cells using a coculture system showed multiple nuclei and expressed albumin protein, similar to characteristics of hepatocytes. Furthermore, these cells expressed the adult hepatocyte marker, CYP3A4. Interestingly, these cells also formed a net structure expressing the bile epithelium capillary marker, multidrug resistance-associated protein 2. Thus, we have demonstrated the induction of a new canine stem cell line, ciXEN-like cells, which could form an embryonic endodermal cell layer. Our ciXEN-like cells may be a helpful tool to study the canine embryo development and represent a promising cell source for proceeding human and canine regenerative medicine.

KEYWORDS:

dog; embryonic stem cell; extraembryonic endoderm cell; hepatocyte; induced extraembryonic endoderm cell; induced pluripotent stem cells

PMID:
28474540
DOI:
10.1089/scd.2017.0026
[Indexed for MEDLINE]

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