Prospective isolation of human bone marrow stromal cell subsets: A comparative study between Stro-1-, CD146- and CD105-enriched populations.
Prospective isolation of human bone marrow stromal cell subsets: A comparative study between Stro-1-, CD146- and CD105-enriched populations.
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人骨髓基质细胞亚群的前瞻性隔离：富含Stro的1-，CD146-和CD105富含种群之间的比较研究。
DOI：
10.1177/2041731414551763
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发表时间：
2014
影响因子：
8.2
通讯作者：
Oreffo RO
中科院分区：
文献类型：
--
作者：
Gothard D;Greenhough J;Ralph E;Oreffo RO
Stro-1已被证明是骨骼干细胞和祖细胞富集的有效标志物，尽管分离的群体仍然是异质的，表现出可变的集落形成效率和成骨分化潜力。骨骼干细胞起源于外膜网状细胞的新发现使两种进一步的标记物脱颖而出，包括CD 146和CD 105（主要是内皮和血管周围）。本研究比较了CD 146-、CD 105-和Stro-1（单独和联合）富集的人骨髓基质细胞亚群，并评估了这些内皮/血管周围标志物是否提供了优于传统Stro-1的进一步选择。荧光细胞分选定量显示，与Stro-1（16.29% ± 0.78%）相比，CD 146和CD 105均靶向较小（分别为2.22% ± 0.59%和6.94% ± 1.34%）且可能不同的人骨髓基质细胞组分。CD 146+，但不是CD 105+，细胞表现出类似的碱性磷酸酶阳性集落形成效率在体外和胶原蛋白/蛋白多糖沉积在体内Stro-1+细胞。对一些选择的成骨和潜在的骨预测基因（包括ALP、CADM 1、CLEC 3B、DCN、LOXL 4、OPN、POL 4和SATB 2）的分子分析显示，Stro-1+和CD 146+群体具有相似的表达谱。分离的人骨髓基质细胞组分（2.04% ± 0.41%）显示Stro-1和CD 146免疫标记阳性。此处提供的数据表明，CD 146+群体与Stro-1+群体相当，但不上级。然而，这项研究表明，迫切需要新的候选标记物，以分离同质的骨骼干细胞群或骨骼干细胞群，表现出同质的体外/体内特性，在组织工程和再生医学策略的实施。
Stro-1 has proved an efficacious marker for enrichment of skeletal stem and progenitor cells although isolated populations remain heterogeneous, exhibiting variable colony-forming efficiency and osteogenic differentiation potential. The emerging findings that skeletal stem cells originate from adventitial reticular cells have brought two further markers to the fore including CD146 and CD105 (both primarily endothelial and perivascular). This study has compared CD146-, CD105- and Stro-1 (individual and in combination)-enriched human bone marrow stromal cell subsets and assessed whether these endothelial/perivascular markers offer further selection over conventional Stro-1. Fluorescent cell sorting quantification showed that CD146 and CD105 both targeted smaller (2.22% ± 0.59% and 6.94% ± 1.34%, respectively) and potentially different human bone marrow stromal cell fractions compared to Stro-1 (16.29% ± 0.78%). CD146+, but not CD105+, cells exhibited similar alkaline phosphatase–positive colony-forming efficiency in vitro and collagen/proteoglycan deposition in vivo to Stro-1+ cells. Molecular analysis of a number of select osteogenic and potential osteo-predictive genes including ALP, CADM1, CLEC3B, DCN, LOXL4, OPN, POSTN and SATB2 showed Stro-1+ and CD146+ populations possessed similar expression profiles. A discrete human bone marrow stromal cell fraction (2.04% ± 0.41%) exhibited positive immuno-labelling for both Stro-1 and CD146. The data presented here show that CD146+ populations are comparable but not superior to Stro-1+ populations. However, this study demonstrates the critical need for new candidate markers with which to isolate homogeneous skeletal stem cell populations or skeletal stem cell populations which exhibit homogeneous in vitro/in vivo characteristics, for implementation within tissue engineering and regenerative medicine strategies.