Adipose-derived stem cells express higher levels of type VII collagen under specific culture conditions
Adipose-derived stem cells express higher levels of type VII collagen under specific culture conditions
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DOI：
10.1007/s00403-017-1781-9
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发表时间：
2017-12-01
影响因子：
3
通讯作者：
Ikeda, Shigaku
中科院分区：
文献类型：
--
作者：
Maeda, Yuichiro;Hasegawa, Toshio;Ikeda, Shigaku
VII型胶原蛋白（Col 7）是真皮表皮连接处锚定原纤维的主要成分。脂肪源性干细胞（ADSC）是一种在再生医学中非常有用的细胞群，因为它们易于分离和具有多向分化的潜力。基于K14在未分化的ADSC中表达并且在分化为脂肪细胞后表达下调的观察结果，我们推测ADSC是角质形成细胞干/祖细胞。在含有全反式维甲酸和骨形态发生蛋白4的培养基中，将脂肪干细胞与成纤维细胞在IV型胶原上共培养。在角质形成细胞无血清培养基中培养的第14天，收获细胞并进行免疫荧光、流式细胞术、实时PCR和蛋白质印迹。大约45%的ADSC对抗人细胞角蛋白10免疫染色阳性，大约80%对Col 7免疫染色阳性。流式细胞术，实时PCR和蛋白质印迹法也证实，分化的脂肪干细胞表达更高水平的Col 7。这些发现支持了ADSC的治疗潜力，不仅用于伤口愈合，而且用于校正Col 7缺陷。
Type VII collagen (Col7) is a major component of the anchoring fibrils at the dermoepidermal junction. Adipose-derived stem cells (ADSCs) are a cell population highly useful in regenerative medicine because of their ease of isolation and their potential for multilineage differentiation. Based on the observations that K14 was expressed in undifferentiated ADSCs and the expression was downregulated after differentiation into adipocytes, we speculated that ADSCs are keratinocyte stem/progenitor cells. ADSCs were co-cultured with fibroblasts on type IV collagen in a medium containing all-trans retinoic acid and bone morphogenetic protein 4. At day 14 of culture in keratinocyte serum-free medium, the cells were harvested and subjected to immunofluorescence, flow cytometry, real-time PCR, and western blotting. Approximately, 45% of ADSCs were immunostained positively for anti-human cytokeratin 10, and approximately 80% were stained positively for Col7. Flow cytometry, real-time PCR, and western blotting also confirmed that differentiated ADSCs expressed higher levels of Col7. These findings support the therapeutic potential of ADSCs, not only for wound healing, but also for the correction of Col7 deficiencies.