Centre for Vaccine Evaluation, Biologics and Genetic Therapies Directorate, Health Products and Food Branch, Health Canada, Ottawa, Canada.
PLoS One. 2012;7(6):e38954. doi: 10.1371/journal.pone.0038954. Epub 2012 Jun 13.
Bone marrow stromal cell cultures contain multipotent cells that may have therapeutic utility for tissue restoration; however, the identity of the cell that maintains this function remains poorly characterized. We have utilized a unique model of murine bone marrow stroma in combination with liquid chromatography mass spectrometry to compare the nuclear, cytoplasmic and membrane associated proteomes of multipotent (MSC) (CD105+) and non-multipotent (CD105-) stromal cells. Among the 25 most reliably identified proteins, 10 were verified by both real-time PCR and Western Blot to be highly enriched, in CD105+ cells and were members of distinct biological pathways and functional networks. Five of these proteins were also identified as potentially expressed in human MSC derived from both standard and serum free human stromal cultures. The quantitative amount of each protein identified in human stromal cells was only minimally affected by media conditions but varied highly between bone marrow donors. This study provides further evidence of heterogeneity among cultured bone marrow stromal cells and identifies potential candidate proteins that may prove useful for identifying and quantifying both murine and human MSC in vitro.
骨髓基质细胞培养物中含有多能细胞,这些细胞可能对组织修复具有治疗作用;然而,维持这种功能的细胞的身份仍未得到很好的描述。我们利用一种独特的鼠骨髓基质模型,结合液相色谱-质谱联用技术,比较了多能(MSC)(CD105+)和非多能(CD105-)基质细胞的核、细胞质和膜相关蛋白质组。在 25 种最可靠鉴定的蛋白质中,有 10 种通过实时 PCR 和 Western Blot 验证,在 CD105+细胞中高度富集,并属于不同的生物学途径和功能网络。其中 5 种蛋白质也被鉴定为可能在来自标准和无血清的人基质培养物中的人 MSC 中表达。在人基质细胞中鉴定的每种蛋白质的定量数量仅受培养基条件的轻微影响,但在骨髓供体之间差异很大。本研究进一步证明了培养的骨髓基质细胞之间存在异质性,并鉴定了潜在的候选蛋白质,这些蛋白质可能有助于鉴定和定量体外的鼠和人 MSC。