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维甲酸和牛磺酸可增强人骨髓基质细胞向视锥光感受器细胞和视网膜神经节细胞的分化。

Retinoic acid and taurine enhance differentiation of the human bone marrow stem cells into cone photoreceptor cells and retinal ganglion cells.

机构信息

Medical Toxicology Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.

Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies, Shahrekord University of Medical Sciences, Shahrekord, Iran.

出版信息

J Cell Biochem. 2021 Dec;122(12):1915-1924. doi: 10.1002/jcb.30151. Epub 2021 Sep 26.

Abstract

Degeneration and apoptotic death of the photoreceptor cell-layer of retina are a major cause of irreversible blindness in the development era. The stem cell replacement therapy is one of the strategies for the retinal repairing. In addition, exogenous signals critically contribute to the direction of lineage decisions that causes the fate-restricted photoreceptor progenitors from stem cell progeny in culture. It has been found that epidermal growth factor (EGF), taurine, and retinoic acid (RA) initially act in the instructive as well as lineage-restricted way in the progenitor lineage for producing neuroretinal cells or photoreceptor like cells from stem cell. The study aims to investigate the effect of RA and taurine in differentiation of the human bone marrow stem cell into cone photoreceptors cells and retinal ganglion cells. Mesenchymal stem cell was derived from human bone marrow of the term delivery. Therefore, the cultured cells have been treated with Dulbecco's modified Eagle's medium (DMEM)/high glucose (H ). After the four-cell passage, basal medium was replaced with DMEM/F12 complemented with 50 μmol/L taurine, RA (1 µM) and EGF (1 µg/ml). Subsequently cellular change morphology was detected following 7 and 14 days. Then, gene expression of neuroretinal and photoreceptor cell biomarkers (CRX, OTX2, PKC-α, recoverin, and Rho) were examined by quantitative polymerase chain reaction (Q-PCR). Also, cells were cultured, fixed, and then immunocytochemical analyzed. Primary antibodies included CRX and Rho. Cellular morphology demonstrated spindle elongated morphology. Taurine alone and combination of RA upregulate neuroretinal and photoreceptor cell biomarkers in messenger RNA and protein levels but along with EGF have not significant effect. Our data showed that taurine combination with RA can differentiate bone marrow mesenchymal stem cells into neuroretinal or photoreceptor like cells in vitro that can offer an attractive treatment ground for transplantation in the cell-replacement therapy for some forms of the retinal degeneration.

摘要

视网膜光感受器细胞层的变性和凋亡是发育时代不可逆失明的主要原因。干细胞替代疗法是视网膜修复的策略之一。此外,外源性信号对谱系决定的方向有重要贡献,导致在培养的干细胞后代中,命运受限的光感受器祖细胞。已经发现表皮生长因子 (EGF)、牛磺酸和视黄酸 (RA) 最初以指导和谱系限制的方式作用于产生神经视网膜细胞或光感受器样细胞的祖细胞谱系从干细胞中。本研究旨在探讨 RA 和牛磺酸对人骨髓干细胞向视锥光感受器细胞和视网膜神经节细胞分化的影响。间充质干细胞来源于足月分娩的人骨髓。因此,培养的细胞用 Dulbecco 修改的 Eagle 培养基 (DMEM)/高葡萄糖 (H) 处理。四细胞传代后,用 DMEM/F12 培养基代替基础培养基,补充 50μmol/L 牛磺酸、RA(1μM)和 EGF(1μg/ml)。随后在第 7 天和第 14 天检测细胞形态变化。然后通过定量聚合酶链反应 (Q-PCR) 检测神经视网膜和光感受器细胞生物标志物 (CRX、OTX2、PKC-α、恢复蛋白和 Rho) 的基因表达。此外,还对细胞进行培养、固定,然后进行免疫细胞化学分析。一抗包括 CRX 和 Rho。细胞形态显示出纺锤形伸长的形态。牛磺酸单独和与 RA 联合上调信使 RNA 和蛋白质水平的神经视网膜和光感受器细胞生物标志物,但与 EGF 联合没有显著影响。我们的数据表明,牛磺酸与 RA 联合可以将骨髓间充质干细胞在体外分化为神经视网膜或光感受器样细胞,这为某些形式的视网膜变性的细胞替代治疗中的移植提供了有吸引力的治疗基础。

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