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人足月胎盘胎儿和母体间充质干/基质细胞系的建立与鉴定

Establishment and characterization of fetal and maternal mesenchymal stem/stromal cell lines from the human term placenta.

作者信息

Qin Sharon Q, Kusuma Gina D, Al-Sowayan Batla, Pace Rishika A, Isenmann Sandra, Pertile Mark D, Gronthos Stan, Abumaree Mohamed H, Brennecke Shaun P, Kalionis Bill

机构信息

University of Melbourne, Department of Obstetrics and Gynaecology, Royal Women's Hospital, Parkville, Victoria, 3052, Australia; Pregnancy Research Centre, Department of Maternal-Fetal Medicine, Royal Women's Hospital, Parkville, Victoria, 3052, Australia.

Mesenchymal Stem Cell Group, Division of Haematology, Institute of Medical and Veterinary Science/Hanson Institute/CSCR, University of Adelaide, Adelaide, SA, 5000, Australia.

出版信息

Placenta. 2016 Mar;39:134-46. doi: 10.1016/j.placenta.2016.01.018. Epub 2016 Jan 22.

Abstract

INTRODUCTION

Human placental mesenchymal stem/stromal cells (MSC) are an attractive source of MSC with great therapeutic potential. However, primary MSC are difficult to study in vitro due to their limited lifespan and patient-to-patient variation.

METHODS

Fetal and maternal MSC were prepared from cells of the chorionic and basal plates of the placenta, respectively. Fetal and maternal MSC were transduced with the human telomerase reverse transcriptase (hTERT). Conventional stem cell assays assessed the MSC characteristics of the cell lines. Functional assays for cell proliferation, cell migration and ability to form colonies in soft agar were used to assess the whether transduced cells retained properties of primary MSC.

RESULTS

Fetal chorionic and maternal MSC were successfully transduced with hTERT to create the cell lines CMSC29 and DMSC23 respectively. The lifespans of CMSC29 and DMSC23 were extended in cell culture. Both cell lines retained important MSC characteristics including cell surface marker expression and multipotent differentiation potential. Neither of the cell lines was tumourigenic in vitro. Gene expression differences were observed between CMSC29 and DMSC23 cells and their corresponding parent, primary MSC. Both cell lines show similar migration potential to their corresponding primary, parent MSC.

DISCUSSION

The data show that transduced MSC retained important functional properties of the primary MSC. There were gene expression and functional differences between cell lines CMSC29 and DMSC23 that reflect their different tissue microenvironments of the parent, primary MSC. CMSC29 and DMSC23 cell lines could be useful tools for optimisation and functional studies of MSC.

摘要

引言

人胎盘间充质干细胞(MSC)是一种具有巨大治疗潜力的有吸引力的MSC来源。然而,原代MSC由于其有限的寿命和个体差异,难以在体外进行研究。

方法

分别从胎盘的绒毛膜和基底板细胞中制备胎儿和母体MSC。用人类端粒酶逆转录酶(hTERT)转导胎儿和母体MSC。常规干细胞检测评估细胞系的MSC特性。采用细胞增殖、细胞迁移和在软琼脂中形成集落能力的功能检测来评估转导细胞是否保留原代MSC的特性。

结果

胎儿绒毛膜和母体MSC分别成功地用hTERT转导,创建了细胞系CMSC29和DMSC23。CMSC29和DMSC23在细胞培养中的寿命延长。两个细胞系都保留了重要的MSC特性,包括细胞表面标志物表达和多能分化潜能。两个细胞系在体外均无致瘤性。观察到CMSC29和DMSC23细胞与其相应的亲本原代MSC之间存在基因表达差异。两个细胞系与其相应的原代亲本MSC表现出相似的迁移潜能。

讨论

数据表明,转导的MSC保留了原代MSC的重要功能特性。细胞系CMSC29和DMSC23之间存在基因表达和功能差异,这反映了它们亲本原代MSC不同的组织微环境。CMSC29和DMSC23细胞系可能是MSC优化和功能研究的有用工具。

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