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Retinoic acid and taurine enhance differentiation of the human bone marrow stem cells into cone photoreceptor cells and retinal ganglion cells
Journal of Cellular Biochemistry ( IF 3.0 ) Pub Date : 2021-09-26 , DOI: 10.1002/jcb.30151 Fatemeh Forouzanfar 1 , Mostafa Soleimannejad 2, 3 , Amin Soltani 2 , Parisa Sadat Mirsafaee 3 , Samira Asgharzade 3, 4
Journal of Cellular Biochemistry ( IF 3.0 ) Pub Date : 2021-09-26 , DOI: 10.1002/jcb.30151 Fatemeh Forouzanfar 1 , Mostafa Soleimannejad 2, 3 , Amin Soltani 2 , Parisa Sadat Mirsafaee 3 , Samira Asgharzade 3, 4
Affiliation
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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.
中文翻译：
维甲酸和牛磺酸促进人骨髓干细胞分化为视锥细胞和视网膜神经节细胞
视网膜感光细胞层的退化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，外源信号对谱系决定的方向做出重要贡献，导致培养中干细胞后代的命运受限的感光祖细胞。已发现表皮生长因子 (EGF)、牛磺酸和视黄酸 (RA) 最初在祖细胞谱系中以指导性和谱系限制的方式起作用，用于从干细胞产生神经视网膜细胞或感光细胞样细胞。本研究旨在探讨RA和牛磺酸在人骨髓干细胞分化为视锥细胞和视网膜神经节细胞中的作用。间充质干细胞来源于人骨髓的术语交付。因此，培养的细胞已经用 Dulbecco 改良 Eagle 培养基 (DMEM)/高糖 (H+）。四细胞传代后，用补充有 50 μmol/L 牛磺酸、RA (1 μM) 和 EGF (1 μg/ml) 的 DMEM/F12 替换基础培养基。随后在 7 天和 14 天后检测到细胞变化形态。然后，通过定量聚合酶链反应 (Q-PCR) 检测神经视网膜和感光细胞生物标志物（CRX、OTX2、PKC-α、recoverin 和 Rho）的基因表达。此外，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。细胞形态显示纺锤形细长形态。单独的牛磺酸和 RA 的组合上调信使 RNA 和蛋白质水平的神经视网膜和感光细胞生物标志物，但与 EGF 一起没有显着影响。
更新日期：2021-09-26
中文翻译：
维甲酸和牛磺酸促进人骨髓干细胞分化为视锥细胞和视网膜神经节细胞
视网膜感光细胞层的退化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，外源信号对谱系决定的方向做出重要贡献，导致培养中干细胞后代的命运受限的感光祖细胞。已发现表皮生长因子 (EGF)、牛磺酸和视黄酸 (RA) 最初在祖细胞谱系中以指导性和谱系限制的方式起作用，用于从干细胞产生神经视网膜细胞或感光细胞样细胞。本研究旨在探讨RA和牛磺酸在人骨髓干细胞分化为视锥细胞和视网膜神经节细胞中的作用。间充质干细胞来源于人骨髓的术语交付。因此，培养的细胞已经用 Dulbecco 改良 Eagle 培养基 (DMEM)/高糖 (H+）。四细胞传代后，用补充有 50 μmol/L 牛磺酸、RA (1 μM) 和 EGF (1 μg/ml) 的 DMEM/F12 替换基础培养基。随后在 7 天和 14 天后检测到细胞变化形态。然后，通过定量聚合酶链反应 (Q-PCR) 检测神经视网膜和感光细胞生物标志物（CRX、OTX2、PKC-α、recoverin 和 Rho）的基因表达。此外，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。细胞形态显示纺锤形细长形态。单独的牛磺酸和 RA 的组合上调信使 RNA 和蛋白质水平的神经视网膜和感光细胞生物标志物，但与 EGF 一起没有显着影响。
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