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Effect of cord blood serum on ex vivo human limbal epithelial cell culture

机译:脐带血血清对离体人角膜缘上皮细胞培养的影响

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摘要

Limbal cell transplantation is an efficacious procedure for rehabilitation of visual acuity in patients with severe ocular surface disorders. Cultivation of limbal epithelial stem cell with fetal bovine serum for transplantation has been a promising treatment for reconstructing the ocular surface in severe limbal stem cell deficiency caused by Steven Johnson syndrome, chemical or thermal injury. This technique of “cell therapy” has been accepted worldwide but the cost of cultivating the cells for transplantation is high. The objective of this study was to investigate the effect of cord blood serum in place of fetal bovine serum on the growth of human limbal epithelial cell culture. Our group has experimented with human cord blood serum which was obtained free of cost from willing donors. The use of human cord blood serum in place of fetal bovine serum for ex vivo culture of limbal stem cell has helped us in reducing the cost of culture. Fresh human limbal tissues from donor cadavers were cultured on intact and denuded amniotic membrane. Cells were proliferated in vitro with cell culture media containing human cord blood serum. Reverse transcription-polymerase chain reaction and immunofluorescence cytochemistry of cultured human limbal epithelial stem cell was done for characterization of the cells.
机译:肢体细胞移植是重度眼表疾病患者视力恢复的有效方法。用胎牛血清培养角膜缘上皮干细胞用于移植已成为在由史蒂文·约翰逊综合征,化学或热损伤引起的严重角膜缘干细胞缺乏症中重建眼表的有前途的治疗方法。这种“细胞疗法”技术已为世界所接受,但培养用于移植的细胞的成本很高。本研究的目的是研究脐带血清代替胎牛血清对人角膜缘上皮细胞培养物生长的影响。我们的小组进行了人脐带血清实验,该血清是从自愿捐助者那里免费获得的。用人脐带血血清代替胎牛血清进行角膜缘干细胞的离体培养有助于我们降低培养成本。在完整且裸露的羊膜上培养来自供体尸体的新鲜人缘组织。细胞在体外用含有人脐带血血清的细胞培养基增殖。进行了培养的人角膜缘上皮干细胞的逆转录-聚合酶链反应和免疫荧光细胞化学的表征。

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