Köllmer Melanie, Buhrman Jason S, Zhang Yu, Gemeinhart Richard A
Department of Biopharmaceutical Sciences, University of Illinois, Chicago, IL 60612-7231, USA.
J Dev Biol Tissue Eng. 2013 May 1;5(2):18-25. doi: 10.5897/JDBTE2013.0065.
The stem cell differentiation paradigm is based on the progression of cells through generations of daughter cells that eventually become restricted and committed to one lineage resulting in fully differentiated cells. Herein, we report on the differentiation of adult human mesenchymal stem cells (hMSCs) towards adipogenic and osteogenic lineages using established protocols. Lineage specific geneswere evaluated by quantitative real-time PCR relative to two reference genes. The expression of osteoblast-associated genes (alkaline phosphatase, osteopontin, and osteocalcin)was detected in hMSCs that underwent adipogenesis. When normalized, the expression of adipocyte marker genes (adiponectin, fatty acid binding protein P4, and leptin) increasedin a time-dependent manner during adipogenic induction. Adiponectin and leptin were also detected in osteoblast-induced cells. Lipid vacuoles that represent the adipocyte phenotype were only present in the adipogenic induction group. Conforming to the heterogeneous nature of hMSCs and the known plasticity between osteogenic and adipogenic lineages, these data indicatea marker overlap between MSC-derived adipocytes and osteoblasts. Weproposea careful consideration of experimental conditions such as investigated timepoints, selected housekeeping genesand the evidence indicating lack of differentiation into other lineageswhen evaluating hMSC differentiation.
干细胞分化模式基于细胞通过子代细胞世代的进展,这些子代细胞最终会受到限制并定向分化为一个谱系,从而产生完全分化的细胞。在此,我们报告了使用既定方案将成人人间充质干细胞(hMSC)向脂肪生成和成骨谱系分化的情况。相对于两个参考基因,通过定量实时PCR评估谱系特异性基因。在经历脂肪生成的hMSC中检测到成骨细胞相关基因(碱性磷酸酶、骨桥蛋白和骨钙素)的表达。标准化后,脂肪细胞标记基因(脂联素、脂肪酸结合蛋白P4和瘦素)的表达在脂肪生成诱导过程中呈时间依赖性增加。在成骨诱导细胞中也检测到了脂联素和瘦素。代表脂肪细胞表型的脂质空泡仅存在于脂肪生成诱导组中。符合hMSC的异质性以及成骨和成脂谱系之间已知的可塑性,这些数据表明源自MSC的脂肪细胞和成骨细胞之间存在标记重叠。我们建议在评估hMSC分化时,仔细考虑实验条件,如研究的时间点、选定的管家基因以及表明未分化为其他谱系的证据。
J Dev Biol Tissue Eng. 2013-5-1
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