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利用保存完好的去细胞大鼠肾脏支架和间充质干细胞进行肾脏组织工程。

Kidney tissue engineering using a well-preserved acellular rat kidney scaffold and mesenchymal stem cells.

作者信息

Shahraki Samira, Moghaddam Matin Maryam, Ebrahimzadeh Bideskan Alireza, Aslzare Mohammad, Bahrami Ahmad Reza, Hosseinian Sara, Iranpour Sonia, Samadi Noshahr Zahra, Khajavi Rad Abolfazl

机构信息

Department of Physiology, School of Medicine, Zahedan University of Medical Sciences, Zahedan, Iran.

Department of Physiology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.

出版信息

Vet Res Forum. 2021;12(3):339-348. doi: 10.30466/vrf.2019.104640.2491. Epub 2021 Sep 15.

Abstract

The aim of this study was to acquire an effective method for preparation of rat decellularized kidney scaffolds capable of supporting proliferation and differentiation of human adipose tissue derived mesenchymal stem cells (AD-MSCs) into kidney cells. We compared two detergents, the sodium dodecyl sulfate (SDS) and triton X-100 for decellularization. The efficiency of these methods was assessed by Hematoxylin and Eosin (H&E), 4', 6 diamidino-2-phenylindole and immunohistochemistry (IHC) staining. In the next step, AD-MSCs were seeded into the SDS-treated scaffolds and assessed after three weeks of culture. Proliferation and differentiation of AD-MSCs into kidney-specific cell types were then analyzed by H&E and IHC staining. The histological examinations revealed that SDS was more efficient in removing kidney cells at all-time points compared to triton X-100. Also, in the SDS-treated sections the native extracellular matrix was more preserved than the triton-treated samples. Laminin was completely preserved during decellularization procedure using SDS. Cell attachment in the renal scaffold was observed after recellularization. Furthermore, differentiation of AD-MSCs into epithelial and endothelial cells was confirmed by expression of Na-K ATPase and vascular endothelial growth factor receptor 2 (VEGFR-2) in seeded rat renal scaffolds, respectively. Our findings illustrated that SDS was more effective for decellularization of rat kidney compared to triton X-100. We presented an optimized method for decellularization and recellularization of rat kidneys to create functional renal natural scaffolds. These natural scaffolds supported the growth of AD-MSCs and could also induce differentiation of these cells into epithelial and endothelial cells.

摘要

本研究的目的是获得一种有效的方法来制备大鼠去细胞肾脏支架,该支架能够支持人脂肪组织来源的间充质干细胞(AD-MSCs)增殖并分化为肾细胞。我们比较了两种用于去细胞处理的去污剂,即十二烷基硫酸钠(SDS)和曲拉通X-100。通过苏木精和伊红(H&E)染色、4',6-二脒基-2-苯基吲哚染色及免疫组织化学(IHC)染色评估这些方法的效率。下一步,将AD-MSCs接种到经SDS处理的支架上,并在培养三周后进行评估。然后通过H&E染色和IHC染色分析AD-MSCs向肾特异性细胞类型的增殖和分化情况。组织学检查显示,与曲拉通X-100相比,SDS在所有时间点去除肾细胞的效率更高。此外,在经SDS处理的切片中,天然细胞外基质比经曲拉通处理的样本保存得更好。在使用SDS进行去细胞处理过程中,层粘连蛋白完全得以保留。再细胞化后观察到细胞在肾支架中的附着情况。此外,分别通过接种大鼠肾支架中Na-K ATP酶和血管内皮生长因子受体2(VEGFR-2)的表达,证实了AD-MSCs向上皮细胞和内皮细胞的分化。我们的研究结果表明,与曲拉通X-100相比,SDS对大鼠肾脏去细胞处理更有效。我们提出了一种优化的大鼠肾脏去细胞和再细胞化方法,以创建功能性肾脏天然支架。这些天然支架支持AD-MSCs的生长,还能诱导这些细胞分化为上皮细胞和内皮细胞。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/02b7/8576151/28c8671f4cc8/vrf-12-339-g001.jpg

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