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Ficoll 密度梯度沉淀法分离成年小鼠背部皮肤的毳毛毛囊间充质干细胞:一种新型的毛囊间充质干细胞分离方法。

Ficoll density gradient sedimentation isolation of pelage hair follicle mesenchymal stem cells from adult mouse back skin: a novel method for hair follicle mesenchymal stem cells isolation.

机构信息

Department of Plastic and Aesthetic Surgery, Nanfang Hospital of Southern Medical University, 1838 Guangzhou North Road, Guangzhou, Guangdong, People's Republic of China.

出版信息

Stem Cell Res Ther. 2022 Jul 28;13(1):372. doi: 10.1186/s13287-022-03051-3.

Abstract

BACKGROUND

Hair follicle mesenchymal stem cells (HF-MSCs) have great potential for cell therapy. Traditional method to isolate whisker HF-MSC is time-consuming and few in cell numbers. How to quickly and conveniently obtain a large number of HF-MSC for experimental research is a problem worth exploring.

METHODS

Two-step Ficoll Density Gradient Sedimentation (FDGS) was performed to isolate pelage HF-MSC from adult mice. The characteristic of the isolated cells was identified and compared with whisker HF-MSC by immunofluorescence staining, flow cytometry, three-lineage differentiation and hair follicle reconstruction. Pelage HF-MSC and exosomes were injected into the dorsal skin of mice as well as hair follicle organ culture to explore its role in promoting hair growth. The cells and exosomes distribution were located by immunofluorescence staining.

RESULTS

Isolated pelage HF-MSC expressed similar markers (ALP, Versican, NCAM, Nestin), showed similar growth pattern, possessed similar mesenchymal stem cells function and hair follicle induction ability as whisker HF-MSC. A large number of cells can be obtained with fewer mice compared to traditional method. Injected pelage HF-MSC promoted hair growth by secreting exosomes.

CONCLUSION

A large number of Pelage HF-MSC can be isolated by FDGS, which can promote hair growth by secreting exosomes which may target the dermal papilla and hair matrix region of host hair follicle.

摘要

背景

毛囊间充质干细胞(HF-MSCs)在细胞治疗中有很大的潜力。传统的分离胡须 HF-MSC 的方法耗时耗力,且细胞数量较少。如何快速方便地获得大量 HF-MSC 用于实验研究是一个值得探索的问题。

方法

采用两步 Ficoll 密度梯度沉降(FDGS)法从成年小鼠的体被毛中分离毛囊间充质干细胞。通过免疫荧光染色、流式细胞术、三系分化和毛囊重建对分离的细胞进行鉴定,并与胡须 HF-MSC 进行比较。将体被毛 HF-MSC 和外泌体注射到小鼠背部皮肤和毛囊器官培养物中,以探索其促进毛发生长的作用。通过免疫荧光染色定位细胞和外泌体的分布。

结果

分离的体被毛 HF-MSC 表达相似的标志物(碱性磷酸酶、软骨素蛋白聚糖、神经细胞黏附分子、巢蛋白),具有相似的生长模式,具有相似的间充质干细胞功能和毛囊诱导能力与胡须 HF-MSC 相同。与传统方法相比,用较少的小鼠可以获得大量的细胞。注射的体被毛 HF-MSC 通过分泌外泌体促进毛发生长。

结论

FDGS 可大量分离体被毛 HF-MSC,通过分泌外泌体促进毛发生长,外泌体可能作用于宿主毛囊的真皮乳头和毛基质区域。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a338/9330686/4a8930cd8670/13287_2022_3051_Fig1_HTML.jpg

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