The assessment of the in vivo to in vitro cellular transition of human umbilical cord multipotent stromal cells
The assessment of the in vivo to in vitro cellular transition of human umbilical cord multipotent stromal cells
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DOI：
10.1016/j.placenta.2014.11.024
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发表时间：
2015-02-01
期刊：
影响因子：
3.8
通讯作者：
Can, A.
中科院分区：
文献类型：
--
作者：
Coskun, H.;Can, A.
简介：人脐带基质是原始多能基质细胞（hUC-MSCs）的丰富来源。然而，hUC-MSC分离和增殖的方法仍然存在争议，并且在实验室之间存在差异。我们的小组先前证明了在酶促分离hUC-MSC后出现两种细胞类型，其随后在以后的传代中经历向成纤维细胞样表型的转变。本研究的目的是通过评估细胞骨架和细胞粘附蛋白并通过比较脐带切片中这些蛋白的重塑来进一步分析培养的hUC-MSCs，以确定由于酶促和外植体方法引起的细胞改变。通过酶或外植体方法分离的组织切片和培养细胞进行形态学分析，并通过标记细胞角蛋白、波形蛋白、α-平滑肌肌动蛋白，E-cadherin和N-cadherin profiles.Results：目前的观察结果证实，宽，扁平的细胞（1型）共享肌纤维母细胞的功能，只出现在酶分离的早期培养物，逐渐减少或取代成纤维细胞样细胞（2型）在以后的通道。相比之下，外植体方法不会导致1型细胞在体外的存在。在测试的CK亚型中，在两种方案后培养时，CK 18表达上调，而CK 19表达下调。波形蛋白和α-SMA，作为主要的中间丝的hUC-间充质干细胞被发现在整个培养期间不变，无论细胞分离technique used.Discussion：所提供的数据证实，并进一步阐明了先前观察到的表型变化，在hUC-间充质干细胞的结构蛋白在酶促分离和随后的培养细胞相比，在原位等价物的改变所示。(C)2014爱思唯尔有限公司版权所有。
Introduction: Human umbilical cord stroma is a rich source of primitive multipotent stromal cells (hUC-MSCs). However, the methods for hUC-MSC isolation and propagation remain controversial and vary among laboratories. Our group previously demonstrated that two cell types emerge upon enzymatic isolation of hUC-MSCs, which subsequently undergo a transition towards a fibroblastoid phenotype in later passages. The aim of this study was to further analyse cultured hUC-MSCs by evaluating the cytoskeletal and cell adhesion proteins and by comparing the remodelling of those proteins in umbilical cord sections to determine the cell alterations due to enzymatic and explant methods.Methods: Tissue sections and cultured cells isolated by enzymatic or explant methods were analysed morphologically and by labelling cytokeratin, vimentin, alpha-smooth muscle actin, E-cadherin and N-cadherin profiles.Results: The present observations confirmed that wide, flat cells (type-1) share myofibroblastic features, appear exclusively in enzymatically isolated early cultures; gradually diminish or are replaced by fibroblastoid cells (type-2) in later passages. In contrast, the explant method does not result in the existence of type-1 cells in vitro. Among the tested CK subtypes, CK18 expression is upregulated, whereas CK19 expression is downregulated upon culturing after both protocols. Vimentin and alpha-SMA, as the major intermediate filaments of hUC-MSCs were found unaltered throughout the culturing period regardless of the cell isolation technique used.Discussion: The data presented confirm and further elucidate the previously observed phenotypic change in hUC-MSCs as illustrated by alterations in structural proteins during enzymatic isolation and subsequent culturing of cells compared with in situ equivalents. (C) 2014 Elsevier Ltd. All rights reserved.