Erythropoietin producing cells for potential cell therapy
Erythropoietin producing cells for potential cell therapy
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DOI：
10.1007/s00345-008-0301-0
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发表时间：
2008-08-01
影响因子：
3.4
通讯作者：
Yoo, James J.
中科院分区：
文献类型：
--
作者：
Aboushwareb, Tamer;Egydio, Fernanda;Yoo, James J.
贫血是慢性肾功能衰竭的必然结果，原因是肾脏产生促红细胞生成素(EPO)的能力降低。我们研究了分离和扩增产生EPO的细胞用于细胞治疗的可行性。材料和方法培养扩增7-10日龄小鼠的肾脏细胞。用免疫细胞化学、流式细胞术和免疫印迹分析，结合EPO特异性抗体，对每一代培养阶段的细胞进行EPO表达的鉴定。结果培养的肾细胞在各传代阶段(P1~P3)均表达EPO，免疫细胞化学分析结果表明，培养的肾细胞表达EPO的能力较强。经洗涤剂溶解的细胞提取液进行Western Blot分析，检测到各代肾细胞中均有EPO(34 KDa)蛋白的表达。结论本实验结果表明，体外培养的肾细胞可产生EPO。该细胞在多个传代阶段稳定表达EPO，并能在体内形成组织。这项研究表明，产生EPO的细胞可能被用作治疗慢性肾功能衰竭引起的贫血的潜在选择。
Introduction Anemia is an inevitable outcome of chronic renal failure due to the kidney's decreased ability to produce erythropoietin (EPO). We examined the feasibility of isolating and expanding EPO-producing cells for cell-based therapy.Materials and methods Renal cells from 7- to 10-day-old mice were culture-expanded. The cells at each subculture stage were characterized for EPO expression, using immunocytochemistry, FACS, and Western Blot analysis, with EPO-specific antibodies. To assess the levels of EPO expression, cells incubated under normoxic and hypoxic conditions were analyzed.Results Immunocytochemical analysis of the cultured renal cells expressed EPO at each subculture stage (P1-P3). Western Blot analysis of the detergent-solubilized cell extracts detected EPO (34 kDa) protein in the kidney cells of all passages tested.Conclusion These results demonstrate that EPO-producing renal cells can be grown and expanded in culture. The cells stably expressed EPO at multiple subculture stages and they are able to form tissue in vivo. This study shows that EPO-producing cells may be used as a potential treatment option for anemia caused by chronic renal failure.