Evaluation of Senescence in Mesenchymal Stem Cells Isolated from Equine Bone Marrow, Adipose Tissue, and Umbilical Cord Tissue
Evaluation of Senescence in Mesenchymal Stem Cells Isolated from Equine Bone Marrow, Adipose Tissue, and Umbilical Cord Tissue
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DOI：
10.1089/scd.2010.0589
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发表时间：
2012-01-01
影响因子：
4
通讯作者：
Borjesson, Dori L.
中科院分区：
文献类型：
--
作者：
Vidal, Martin A.;Walker, Naomi J.;Borjesson, Dori L.
来自成人和新生儿组织的间充质干细胞（MSC）被广泛研究用于再生医学。本研究的目的是比较从马骨髓（BMSC）、脂肪组织（ASC）和脐带组织（UCMSC）分离的MSC的复制性衰老的开始。评估了从4个不同供体组织中收获的MSC的MSC增殖（细胞倍增）、衰老相关的β-半乳糖苷酶染色、端粒长度、Sox-2和谱系特异性标志物表达。结果显示，在随后的衰老之前，所有细胞类型都以类似于1天/细胞倍增的速度增殖。BMSC在第10代时显著增加群体倍增率，并且在略大于30个总群体倍增后停止增殖，而UCMSC和ASCs达到约60至80个总群体倍增。UCMSC和ASCs在类似于70个群体的双染后显示明显的b-半乳糖苷酶染色，而BMSCs在类似于30个群体的双染后显示阳性。衰老的发生与端粒长度的显著减少相关，在第3代平均为10.2 kbp，在衰老培养物中为4.5 kbp。与早期传代相比，MSC在衰老时骨粘连蛋白染色强烈，而波形蛋白和低水平的平滑肌肌动蛋白表达一致。Sox-2基因表达在所有3种MSC类型中一致。总之，马BMSC似乎比ASC和UCMSC衰老得早得多。这些结果表明可用于研究和组织工程的传代培养BMSC的传代次数有限，并表明脂肪组织和脐带组织可能更适合用于组织库目的。
Mesenchymal stem cells (MSCs) from adult and neonatal tissues are intensively investigated for their use in regenerative medicine. The purpose of this study was to compare the onset of replicative senescence in MSCs isolated from equine bone marrow (BMSC), adipose tissue (ASC), and umbilical cord tissue (UCMSC). MSC proliferation (cell doubling), senescence-associated beta-galactosidase staining, telomere length, Sox-2, and lineage-specific marker expression were assessed for MSCs harvested from tissues of 4 different donors. The results show that before senescence ensued, all cell types proliferated at similar to 1 day/cell doubling. BMSCs significantly increased population doubling rate by passage 10 and ceased proliferation after a little >30 total population doublings, whereas UCMSCs and ASCs achieved about 60 to 80 total population doublings. UCMSC and ASCs showed marked b-galactosidase staining after similar to 70 population doublings, whereas BMSCs stained positive by similar to 30 population doublings. The onset of senescence was associated with a significant reduction in telomere length averaging 10.2 kbp at passage 3 and 4.5 kbp in senescent cultures. MSCs stained intensively for osteonectin at senescence compared with earlier passages, whereas vimentin and low levels of smooth muscle actin were consistently expressed. Sox-2 gene expression was consistently noted in all 3 MSC types. In conclusion, equine BMSCs appear to senesce much earlier than ASCs and UCMSCs. These results demonstrate the limited passage numbers of subcultured BMSCs available for use in research and tissue engineering and suggest that adipose tissue and umbilical cord tissue may be preferable for tissue banking purposes.