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人毛囊干细胞在转化生长因子-β1 和血小板衍生生长因子-BB 的诱导下分化为收缩性平滑肌细胞。

Human hair follicle stem cell differentiation into contractile smooth muscle cells is induced by transforming growth factor-β1 and platelet-derived growth factor BB.

机构信息

Department of Plastic and Reconstructive Surgery, Shanghai 9th People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200011, P.R. China.

出版信息

Mol Med Rep. 2013 Dec;8(6):1715-21. doi: 10.3892/mmr.2013.1707. Epub 2013 Sep 27.

Abstract

Smooth muscle cells (SMCs) are important in vascular homeostasis and disease and thus, are critical elements in vascular tissue engineering. Although adult SMCs have been used as seed cells, such mature differentiated cells suffer from limited proliferation potential and cultural senescence, particularly when originating from older donors. By comparison, human hair follicle stem cells (hHFSCs) are a reliable source of stem cells with multi-differentiation potential. The aim of the present study, was to develop an efficient strategy to derive functional SMCs from hHFSCs. hHFSCs were obtained from scalp tissues of healthy adult patients undergoing cosmetic plastic surgery. The hHFSCs were expanded to passage 2 and induced by the administration of transforming growth factor-β1 (TGF-β1) and platelet-derived growth factor BB (PDGF-BB) in combination with culture medium. Expression levels of SMC-related markers, including α-smooth muscle actin (α-SMA), α-calponin and smooth muscle myosin heavy chain (SM-MHC), were detected by immunofluorescence staining, flow cytometry analysis and reverse transcription-polymerase chain reaction (RT-PCR). When exposed to differentiation medium, hHFSCs expressed early, mid and late markers (α-SMA, α-calponin and SM-MHC, respectively) that were similar to the markers expressed by human umbilical artery SMCs. Notably, when entrapped inside a collagen matrix lattice, these SM differentiated cells showed a contractile function. Therefore, the present study developed an efficient strategy for differentiating hHFSCs into contractile SMCs by stimulation with TGF-β1 and PDGF-BB. The high yield of derivation suggests that this strategy facilitates the acquisition of the large numbers of cells that are required for blood vessel engineering and the study of vascular disease pathophysiology.

摘要

平滑肌细胞(SMCs)在血管稳态和疾病中起着重要作用,因此是血管组织工程的关键元素。尽管成年 SMC 已被用作种子细胞,但这些成熟的分化细胞增殖潜力有限,且存在培养衰老问题,尤其是来源于老年供体时。相比之下,人毛囊干细胞(hHFSCs)是一种具有多向分化潜能的可靠干细胞来源。本研究旨在开发一种有效的策略,从 hHFSCs 中获得功能性 SMC。hHFSCs 从接受美容整形手术的健康成年患者的头皮组织中获得。将 hHFSCs 扩增至第 2 代,然后通过给予转化生长因子-β1(TGF-β1)和血小板衍生生长因子 BB(PDGF-BB)与培养基联合诱导。通过免疫荧光染色、流式细胞术分析和逆转录-聚合酶链反应(RT-PCR)检测 SMC 相关标志物的表达水平,包括α-平滑肌肌动蛋白(α-SMA)、α-钙调蛋白和平滑肌肌球蛋白重链(SM-MHC)。当暴露于分化培养基时,hHFSCs 表达了早期、中期和晚期标志物(分别为α-SMA、α-钙调蛋白和 SM-MHC),与人脐动脉 SMCs 表达的标志物相似。值得注意的是,当这些分化的 SM 细胞被包裹在胶原基质格子中时,它们表现出收缩功能。因此,本研究通过刺激 TGF-β1 和 PDGF-BB 开发了一种将 hHFSCs 分化为收缩性 SMC 的有效策略。高产量的衍生表明,这种策略有助于获得大量用于血管工程和血管疾病病理生理学研究所需的细胞。

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