Department of Stem Cells and Developmental Biology, Cell Sciences Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran; Department of Biology, Roudehen Branch, Islamic Azad University, Roudehen, Iran.
Department of Regenerative Medicine, Cell Sciences Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.
Differentiation. 2019 Sep-Oct;109:1-8. doi: 10.1016/j.diff.2019.07.001. Epub 2019 Jul 10.
End-stage renal disease (ESRD) is a major global public health issue. In the past decade, regenerative medicine and cell-based therapies were recommended for treatment of devastating diseases like ESRD. Renal progenitor (RP) cells are essential players in such treatment approaches. The major practical difficulties in application of RP cells are generation of these cells and preservation of their self-renewal capacity; also, they should lack identified appropriate cell surface markers. To identify and isolate RP cells, two cell surface markers namely, CD133 and CD24 were recently used. In this study, we used these markers to facilitate selection and purification of RP cells from embryoid bodies (EBs), and assessed the impact of the use of bFGF on frequency of CD133CD24 expression in cells presented in EBs. Moreover, following isolation of CD133CD24 cells from EBs, we evaluated the effect of embryonic, neonatal and adult mouse kidney-derived mesenchymal stem cells (E-KMSC, N-KMSC and A-KMSC respectively) and fibronectin on further differentiation of the sorted cells. Hence, we cultured undifferentiated human embryonic stem cells (hESCs) in suspension state in the presence or absence of bFGF and determined maximum number of CD133CD24 cells in bFGF-treated EBs on day 7. Then, we tested the effect of E-KMSC co-culture and seeding on fibronectin-coated plated on differentiation of the sorted cells into renal epithelial cells. Results revealed down-regulation of several RP cells, markers in CD133CD24 cells. In contrast, renal epithelial marker gene expressions were up-regulated after 7 days of co-culture with E-KMSC. Furthermore, fibronectin resulted in higher expression of renal epithelial markers compared to the E-KMSC co-cultured cells. All in all, bFGF could enhance the number of RP cells expressing CD133 and CD24 markers, in human EBs. We suggest E-KMSC and fibronectin as a promising supplementary factor to further induce differentiation of RP cells into renal epithelial cells.
终末期肾病（ESRD）是一个主要的全球公共卫生问题。在过去的十年中，再生医学和基于细胞的疗法被推荐用于治疗 ESRD 等破坏性疾病。肾祖细胞（RP）细胞是这些治疗方法的重要参与者。应用 RP 细胞的主要实际困难是这些细胞的产生和自我更新能力的保持；此外，它们应该缺乏已确定的适当的细胞表面标记物。为了鉴定和分离 RP 细胞，最近使用了两种细胞表面标记物，即 CD133 和 CD24。在这项研究中，我们使用这些标记物来促进从胚状体（EB）中选择和纯化 RP 细胞，并评估 bFGF 的使用对 EB 中细胞 CD133CD24 表达频率的影响。此外，在从 EB 中分离出 CD133CD24 细胞后，我们评估了胚胎、新生和成年小鼠肾脏来源的间充质干细胞（E-KMSC、N-KMSC 和 A-KMSC 分别）和纤连蛋白对分选细胞进一步分化的影响。因此，我们在存在或不存在 bFGF 的情况下将未分化的人胚胎干细胞（hESC）悬浮培养，并在第 7 天测定 bFGF 处理的 EB 中 CD133CD24 细胞的最大数量。然后，我们测试了 E-KMSC 共培养和接种到纤连蛋白包被的平板上对分选细胞分化为肾上皮细胞的影响。结果显示，CD133CD24 细胞中的几个 RP 细胞标记物表达下调。相反，与 E-KMSC 共培养 7 天后，肾上皮标记基因的表达上调。此外，与 E-KMSC 共培养的细胞相比，纤连蛋白导致肾上皮标记物的表达更高。总之，bFGF 可以增加在人 EB 中表达 CD133 和 CD24 标记物的 RP 细胞数量。我们建议 E-KMSC 和纤连蛋白作为进一步诱导 RP 细胞分化为肾上皮细胞的有前途的补充因子。
