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人羊膜间充质干细胞的分离与鉴定

Isolation and characterization of mesenchymal stem cells from human amniotic membrane.

作者信息

Díaz-Prado Silvia, Muiños-López Emma, Hermida-Gómez Tamara, Rendal-Vázquez Ma Esther, Fuentes-Boquete Isaac, de Toro Francisco J, Blanco Francisco J

机构信息

1 Department of Medicine, INIBIC-University of A Coruña , A Coruña, Spain .

出版信息

Tissue Eng Part C Methods. 2011 Jan;17(1):49-59. doi: 10.1089/ten.TEC.2010.0136. Epub 2010 Aug 30.

Abstract

INTRODUCTION

The human amniotic membrane is a highly abundant and readily available tissue that may be useful for regenerative medicine and cell therapy.

AIM

To compare two previously published protocols for the isolation of human amnion mesenchymal stromal cells (hAMSCs), including their phenotypic characterization and in vitro potential for differentiation toward osteogenic, adipogenic, and chondrogenic mesodermal lineages.

MATERIALS AND METHODS

Human placentas were obtained from selected caesarean-sectioned births. Two different protocols (Alviano et al. (1) and Soncini et al. (2) ) for the isolation of hAMSCs were performed. After monolayer expansion of adherent cells from both protocols, the cells were characterized by flow cytometry and for multipotentiality, as assessed by their capability to differentiate toward adipocyte-, osteoblast-, and chondrocyte-like cells.

RESULTS

Both protocols yielded hAMSCs that showed plastic adherence, fibroblast-like growth, and well-defined human MSC markers. The cell yield and mesodermal differentiation capability of hAMSCs were higher in cells isolated using the Soncini protocol.

CONCLUSIONS

Our data demonstrated the successful isolation of hAMSCs from full-term placentas using two published protocols. Differences between the two protocols in cell yield and in vitro differentiation potential are shown.

摘要

引言

人羊膜是一种非常丰富且易于获取的组织,可能对再生医学和细胞治疗有用。

目的

比较两种先前发表的分离人羊膜间充质基质细胞(hAMSCs)的方案,包括其表型特征以及体外向成骨、成脂和软骨生成中胚层谱系分化的潜力。

材料与方法

从选定的剖宫产分娩中获取人胎盘。执行两种不同的分离hAMSCs的方案(Alviano等人(1)和Soncini等人(2))。在对两种方案中贴壁细胞进行单层扩增后,通过流式细胞术对细胞进行表征,并通过其向脂肪细胞样、成骨细胞样和软骨细胞样细胞分化的能力评估其多能性。

结果

两种方案均产生了表现出塑料贴壁、成纤维细胞样生长以及明确的人MSC标志物的hAMSCs。使用Soncini方案分离的细胞中,hAMSCs的细胞产量和中胚层分化能力更高。

结论

我们的数据表明使用两种已发表的方案成功地从足月胎盘中分离出了hAMSCs。显示了两种方案在细胞产量和体外分化潜力方面的差异。

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