Human skeletal muscle fibroblasts, but not myogenic cells, readily undergo adipogenic differentiation
Human skeletal muscle fibroblasts, but not myogenic cells, readily undergo adipogenic differentiation
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DOI：
10.1242/jcs.132563
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发表时间：
2013-12-15
影响因子：
4
通讯作者：
Harridge, Stephen D. R.
中科院分区：
文献类型：
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作者：
Agley, Chibeza C.;Rowlerson, Anthea M.;Harridge, Stephen D. R.
我们对通过酶消化从人骨骼肌中分离的贴壁细胞类型进行了表征，并证明即使在分离后72小时，这些培养物也主要由肌源性细胞（CD 56（+）、结蛋白（+））和成纤维细胞（TE-7（+）、胶原VI+、PDGFR α（+）、波形蛋白（+）、纤连蛋白（+））组成。为了评估获得的细胞类型的行为，我们优化了一种双重免疫磁性细胞分选方法，用于从成纤维细胞中分离肌源性细胞。该方法得到>96%的肌原性（CD 56（+）结蛋白（+））细胞纯度。从第一次分选获得的CD 56（-）级分高度富集在TE-7（+）成纤维细胞中。使用定量分析的脂质含量，谱系标志物和转录因子的免疫荧光染色，我们测试了纯化的细胞群是否可以分化成脂肪细胞在处理与脂肪酸或脂肪细胞诱导培养基。两种处理均导致成纤维细胞分化为脂肪细胞，如细胞内TE-7的丢失、成脂转录因子PPAR γ和C/EBP α的上调以及采用脂质负载的脂肪细胞形态所示。相比之下，成肌细胞不经历脂肪形成，并显示出对这些脂肪形成处理的响应的PPAR γ和C/EBP α的差异调节。我们的研究结果表明，人骨骼肌成纤维细胞至少是双能祖细胞，可以保持为细胞外基质生产细胞或分化为脂肪细胞。
We characterised the adherent cell types isolated from human skeletal muscle by enzymatic digestion, and demonstrated that even at 72 hours after isolation these cultures consisted predominantly of myogenic cells (CD56(+), desmin(+)) and fibroblasts (TE-7(+), collagen VI+, PDGFR alpha(+), vimentin(+), fibronectin(+)). To evaluate the behaviour of the cell types obtained, we optimised a double immuno-magnetic cell-sorting method for the separation of myogenic cells from fibroblasts. This procedure gave purities of >96% for myogenic (CD56(+) desmin(+)) cells. The CD56(-) fraction obtained from the first sort was highly enriched in TE-7(+) fibroblasts. Using quantitative analysis of immunofluorescent staining for lipid content, lineage markers and transcription factors, we tested if the purified cell populations could differentiate into adipocytes in response to treatment with either fatty acids or adipocyte-inducing medium. Both treatments caused the fibroblasts to differentiate into adipocytes, as shown by loss of intracellular TE-7, upregulation of the adipogenic transcription factors PPAR gamma and C/EBP alpha, and adoption of a lipid-laden adipocyte morphology. By contrast, myogenic cells did not undergo adipogenesis and showed differential regulation of PPAR gamma and C/EBP alpha in response to these adipogenic treatments. Our results show that human skeletal muscle fibroblasts are at least bipotent progenitors that can remain as extracellular-matrix-producing cells or differentiate into adipocytes.