Generation of Canine Induced Extraembryonic Endoderm-Like Cell Line That Forms Both Extraembryonic and Embryonic Endoderm Derivatives
Generation of Canine Induced Extraembryonic Endoderm-Like Cell Line That Forms Both Extraembryonic and Embryonic Endoderm Derivatives
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DOI：
10.1089/scd.2017.0026
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发表时间：
2017-08-01
影响因子：
4
通讯作者：
Inaba, Toshio
中科院分区：
文献类型：
--
作者：
Nishimura, Toshiya;Unezaki, Naoya;Inaba, Toshio
胚外内胚层（extraembryonic endoderm，XEN）细胞是来源于胚泡内细胞团的原始内胚层细胞的干细胞系。这些细胞具有自我更新的特性，并分化成卵黄囊的内脏内胚层（VE）和壁内胚层（PE）。最近，有报道称，XEN细胞可以促进胎儿胚胎内胚层，并对其独特的效力进行了评估。在这项研究中，我们描述了新的犬干细胞系，非常类似于XEN细胞的诱导和表征。这些细胞，我们命名为犬诱导XEN（ciXEN）样细胞，诱导犬胚胎成纤维细胞通过引入四个转基因。CiXEN样细胞表达XEN标记物，其可以在补充有丝分裂原活化蛋白激酶p38和转化生长因子β 1的抑制剂的N2B27培养基中维持超过50代。我们的ciXEN样细胞在没有转基因表达的情况下维持，并且在无饲养层条件下表现出VE和PE标志物的上调表达。使用共培养系统从ciXEN样细胞分化的细胞显示多个核并表达白蛋白，类似于肝细胞的特征。此外，这些细胞表达成人肝细胞标志物CYP 3A4。有趣的是，这些细胞还形成了表达胆汁上皮毛细血管标记物多药耐药相关蛋白2的网状结构。因此，我们已经证明了一种新的犬干细胞系，ciXEN样细胞，它可以形成胚胎内胚层细胞层的诱导。我们的ciXEN样细胞可能是一个有用的工具，研究犬胚胎发育，并代表了一个有前途的细胞来源，进行人类和犬再生医学。
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.