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用于神经组织工程应用的电纺纳米纤维支架上人类头皮脂肪间充质干细胞向成熟神经细胞的分化

Differentiation of Human Scalp Adipose-Derived Mesenchymal Stem Cells into Mature Neural Cells on Electrospun Nanofibrous Scaffolds for Nerve Tissue Engineering Applications.

作者信息

Fesharaki Mehrafarin, Razavi Shahnaz, Ghasemi-Mobarakeh Laleh, Behjati Mohaddeseh, Yarahmadian Reyhaneh, Kazemi Mohammad, Hejazi Hossein

机构信息

Department of Cell Sciences Research Center Medical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.

Department of Anatomical Sciences, Medicine School, Isfahan University of Medical Sciences, Isfahan, Iran.

出版信息

Cell J. 2018 Jul;20(2):168-176. doi: 10.22074/cellj.2018.4898. Epub 2018 Mar 18.

Abstract

OBJECTIVES

This study aimed to isolate and culture SADS cells, investigate their neurogenic capacity and evaluate their application for nerve tissue engineering.

MATERIALS AND METHODS

In this experimental study, SADS cells were isolated from human adipose tissue. After 7-day treatment of SADS cells with insulin, indomethacin and isobutylmethylxanthine, neurogenic differentiation of SADS cells was investigated. During this study, Poly (ε-caprolactone) (PCL) and PCL/gelatin nanofibrous scaffolds were fabricated using electrospinning and subsequently nanofibrous scaffolds were coated with platelet-rich plasma (PRP). SADS cells were also seeded on nanofibrous scaffolds and neurogentic differentiation of these cells on nanofibers was also evaluated. Effect of PRP on proliferation and differentiation of SADS cells on scaffolds was also studied.

RESULTS

Our results showed that after 7-day treatment of SADS cells with insulin, indomethacin and isobutylmethylxanthine, SADS cells expressed markers characteristic of neural cells such as nestin and neuron specific nuclear protein (NEUN) (as early neuronal markers) as well as microtubule-associated protein 2 (MAP2) and neuronal microtubule-associated (TAU) (as mature neuronal markers) while mature astrocyte maker (GFAP) was not expressed. MTT assay and SEM results showed that incorporation of gelatin and PRP into the structure of nanofibrous scaffolds has a significant positive influence on the bioactivity of scaffolds. Our results also showed neurogentic differentiation of SADS cells on scaffolds.

CONCLUSIONS

Our results demonstrated that SADS cells have potential to differentiate into early and mature progenitor neurons, in vitro. PCL/gelatin/PRP was found to be a promising substrate for proliferation of SADS cells and differentiation of these cells into neural cells which make these scaffolds a candidate for further in vivo experiments and suggest their application for nerve tissue engineering.

摘要

目的

本研究旨在分离培养脂肪来源干细胞(SADS细胞),研究其神经分化能力,并评估其在神经组织工程中的应用。

材料与方法

在本实验研究中,从人脂肪组织中分离出SADS细胞。在用胰岛素、消炎痛和异丁基甲基黄嘌呤对SADS细胞进行7天处理后,研究SADS细胞的神经分化情况。在本研究过程中,采用静电纺丝法制备聚(ε-己内酯)(PCL)和PCL/明胶纳米纤维支架,随后用富含血小板血浆(PRP)对纳米纤维支架进行涂层处理。将SADS细胞接种在纳米纤维支架上,并评估这些细胞在纳米纤维上的神经分化情况。还研究了PRP对支架上SADS细胞增殖和分化的影响。

结果

我们的结果表明,在用胰岛素、消炎痛和异丁基甲基黄嘌呤对SADS细胞进行7天处理后,SADS细胞表达神经细胞特征性标志物,如巢蛋白和神经元特异性核蛋白(NEUN)(作为早期神经元标志物)以及微管相关蛋白2(MAP2)和神经元微管相关蛋白(TAU)(作为成熟神经元标志物),而成熟星形胶质细胞标志物(GFAP)未表达。MTT法和扫描电镜结果表明,将明胶和PRP掺入纳米纤维支架结构中对支架的生物活性有显著的积极影响。我们的结果还表明SADS细胞在支架上发生了神经分化。

结论

我们的结果表明,SADS细胞在体外有分化为早期和成熟祖神经元的潜力。发现PCL/明胶/PRP是SADS细胞增殖并分化为神经细胞的有前景的基质,这使得这些支架成为进一步体内实验的候选者,并表明它们可应用于神经组织工程。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ba8d/5893287/04e690d9b512/Cell-J-20-168-g01.jpg

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