Clonal analysis and hierarchy of human bone marrow mesenchymal stem and progenitor cells.
Clonal analysis and hierarchy of human bone marrow mesenchymal stem and progenitor cells.
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DOI：
10.1016/j.exphem.2009.11.001
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发表时间：
2010-01
影响因子：
2.6
通讯作者：
Tarantal, Alice F.
中科院分区：
文献类型：
--
作者：
Lee, C. Chang I.;Christensen, Jared E.;Yoder, Mervin C.;Tarantal, Alice F.
本研究旨在评估单个细胞水平的成人骨髓间充质干/祖细胞，并根据增殖潜力确定分级。将表达增强型绿色荧光蛋白（EGFP）的成人骨髓间充质细胞作为单细胞分选到24孔板中，通过荧光显微镜检查用单个EGFP阳性细胞确认每个孔，并且每三天计数一次。扩增来源于单细胞的集落，然后使用已建立的分化方案分选和评价脂肪形成、软骨形成和成骨谱系。通过实时RT-PCR（PPARγ2、LEP、LPL、LUM、COMP、BIG、CBFA 1、IBSP、BGln）和特异于三系分化的免疫细胞化学（PPARγ1/2、胶原蛋白II、骨唾液蛋白II）进一步分析细胞。骨髓间充质细胞包括高增殖潜能间充质集落形成细胞（HPP-MCFC，7%）、低增殖潜能间充质集落形成细胞（LPP-MCFC，29%）、间充质细胞团（MCC，26%）和成熟间充质细胞（MMC，38%）。所有LPP-MCFC、MCC和MMC菌落在评价期结束时均达到衰老。然而，HPP-MCFC继续生长，显示出向所有三个谱系的分化，并证明了在克隆水平上重新铺板后产生次级HPP-MCFC的能力。这些发现表明，存在低频率的骨髓来源的HPP-MCFC，其可以在单细胞水平上自我更新并向间充质来源的多个谱系分化。
This study was performed to assess adult human bone marrow mesenchymal stem/progenitor cells at a single cell level and to determine a hierarchy based on proliferative potential. Adult bone marrow mesenchymal cells expressing the enhanced green fluorescent protein (EGFP) were sorted as single cells into 24-well plates, each well confirmed with single EGFP-positive cells by fluorescence microscopy, and counted every three days. Colonies derived from single cells were expanded then sorted and evaluated using established differentiation protocols for adipogenic, chondrogenic, and osteogenic lineages. Cells were further analyzed by real-time RT-PCR (PPARγ2, LEP, LPL, LUM, COMP, BIG, CBFA1, IBSP, BGLAP) and immunocytochemistry (PPARγ1/2, Collagen II, Bone Sialoprotein II) specific for tri-lineage differentiation. Bone marrow mesenchymal cells were found to contain high proliferative potential-mesenchymal colony-forming cells (HPP-MCFC, 7%), low proliferative potential-mesenchymal colony-forming cells (LPP-MCFC, 29%), mesenchymal cell clusters (MCC, 26%), and mature mesenchymal cells (MMC, 38%). All LPP-MCFC, MCC, and MMC colonies reached senescence at the end of the evaluation period. However, HPP-MCFC continued to grow, showed differentiation toward all three lineages, and demonstrated the capacity to give rise to secondary HPP-MCFC upon replating at a clonal level. These findings suggest that there is a low frequency of bone marrow derived HPP-MCFC that can both self-renew at a single cell level and differentiate toward multiple lineages of mesenchymal origin.
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影响因子：
4
作者：
Hacia, Joseph G.;Lee, C. Chang I.;Tarantal, Alice F.
通讯作者：
Tarantal, Alice F.
影响因子：
64.5
作者：
Sacchetti, Benedetto;Funari, Alessia;Bianco, Paolo
通讯作者：
Bianco, Paolo
影响因子：
--
作者：
Jones, Elena A.;Crawford, Aileen;McGonagle, Dennis
通讯作者：
McGonagle, Dennis
影响因子：
4
作者：
Kim, Brandon S.;Lee, C. Chang I.;Tarantal, Alice F.
通讯作者：
Tarantal, Alice F.
影响因子：
--
作者：
Mareddy, Shobha;Crawford, Ross;Xiao, Yin
通讯作者：
Xiao, Yin