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利用静电纺丝聚乙烯醇/人血清白蛋白/明胶纤维构建人工雄性生殖细胞龛

Development of An Artificial Male Germ Cell Niche Using Electrospun Poly Vinyl Alcohol/Human Serum Albumin/Gelatin Fibers.

作者信息

Borzouie Zahra, Naghibzadeh Majid, Talebi Ali Reza, Pourrajab Fatemeh, Jebali Ali, Nikukar Habib, Molla Hoseini Hosein, Khoradmehr Arezoo, Khoradmehr Arezoo, Sadeghian-Nodoushan Fatemeh, Aflatoonian Behrouz, Hekmatimoghaddam Seyedhossein

机构信息

Stem Cell Biology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.

Research and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.

出版信息

Cell J. 2019 Oct;21(3):300-306. doi: 10.22074/cellj.2019.6120. Epub 2019 Jun 15.

Abstract

OBJECTIVE

Recent achievements in stem cell biotechnology, nanotechnology and tissue engineering have led to development of novel approaches in regenerative medicine. Azoospermia is one of the challenging disorders of the reproductive system. Several efforts were made for isolation and culture of testis-derived stem cells to treat male infertility. However, tissue engineering is the best approach to mimic the three dimensional microenvironment of the testis in vitro. We investigated whether human testis-derived cells (hTCs) obtained by testicular sperm extraction (TESE) can be cultured on a homemade scaffold composed of electrospun nanofibers of homogeneous poly (vinyl alcohol)/human serum albumin/gelatin (PVA/HSA/gelatin).

MATERIALS AND METHODS

In this experimental lab study, human TCs underwent two steps of enzymatic cell isolation and five culture passages. Nanofibrous scaffolds were characterized by scanning electron microscopy (SEM) and Fouriertransform infrared spectroscopy (FTIR). Attachment of cells onto the scaffold was shown by hematoxylin and eosin (H and E) staining and SEM. Cell viability study using MTT [3-(4, 5-dimethyl-2-thiazolyl) -2, 5-diphenyl -2H- tetrazolium bromide] assay was performed on days 7 and 14.

RESULTS

Visualization by H and E staining and SEM indicated that hTCs were seeded on the scaffold. MTT test showed that the PVA/HSA/gelatin scaffold is not toxic for hTCs.

CONCLUSION

It seems that this PVA/HSA/gelatin scaffold is supportive for growth of hTCs.

摘要

目的

干细胞生物技术、纳米技术和组织工程学的最新进展促使再生医学领域出现了新方法。无精子症是生殖系统中具有挑战性的疾病之一。人们为分离和培养睾丸来源的干细胞以治疗男性不育症付出了诸多努力。然而,组织工程是在体外模拟睾丸三维微环境的最佳方法。我们研究了通过睾丸精子提取(TESE)获得的人睾丸来源细胞(hTCs)能否在由均匀的聚(乙烯醇)/人血清白蛋白/明胶(PVA/HSA/明胶)电纺纳米纤维组成的自制支架上培养。

材料与方法

在这项实验性实验室研究中,人睾丸细胞经历了两步酶促细胞分离和五次传代培养。通过扫描电子显微镜(SEM)和傅里叶变换红外光谱(FTIR)对纳米纤维支架进行表征。通过苏木精和伊红(H&E)染色和SEM显示细胞附着在支架上。在第7天和第14天使用MTT [3 -(4,5 - 二甲基 - 2 - 噻唑基)- 2,5 - 二苯基 - 2H - 四唑溴盐] 法进行细胞活力研究。

结果

H&E染色和SEM观察表明hTCs接种在支架上。MTT试验表明PVA/HSA/明胶支架对hTCs无毒。

结论

这种PVA/HSA/明胶支架似乎对hTCs的生长具有支持作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/477e/6582427/1a9b02a5854c/Cell-J-21-300-g01.jpg

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