Sudha B, Madhavan H N, Sitalakshmi G, Malathi J, Krishnakumar S, Mori Y, Yoshioka H, Abraham S
Vision Research Foundation, Sankara Nethralaya, Chennai, India.
Indian J Med Res. 2006 Dec;124(6):655-64.
BACKGROUND & OBJECTIVES: Cultivated limbal stem cell transplantation is being used as a current treatment modality for limbal stem cell deficiency. However, use of allogenic biological material as substrate is associated with risks of transmission of certain diseases and allograft rejection. Therefore development of non-toxic biodegradable synthetic polymers is important. We undertook this study to evaluate the use of a synthetic polymer Mebiol gel as a substrate for the growth of limbal phenotype cells and cornea phenotype cells from limbal explants.
Human cadaveric limbal explants cells were cultivated on Mebiol gel. The proliferative capacity of cultivated cells was analyzed with thymidine incorporation studies. Immunostaining for presumed limbal stem cell association markers and cornea differentiation markers was performed and confirmed with reverse transcription (RT-PCR).
The limbal explants underwent proliferation in vitro. The cultivated cells expressed the presumed limbal stem cell association markers (ABCG2 and p63), the transient amplifying cell markers (connexin 43, integrin alpha9) and the cornea differentiation marker (K3). RT PCR confirmed the immunohistochemical data.
INTERPRETATION & CONCLUSION: Our findings showed that the synthetic polymer Mebiol gel was able to support limbal explant proliferation. The cultured cells expressed presumed limbal stem cell association markers, transient amplifying cells and cornea phenotype markers. Mebiol Gel can be used as a scaffold for growing limbal explants.
培养的角膜缘干细胞移植目前正被用作治疗角膜缘干细胞缺乏症的一种方式。然而，使用同种异体生物材料作为基质存在某些疾病传播和同种异体移植排斥的风险。因此，开发无毒的可生物降解合成聚合物很重要。我们进行了这项研究，以评估合成聚合物Mebiol凝胶作为角膜缘外植体中角膜缘表型细胞和角膜表型细胞生长基质的用途。
将人尸体角膜缘外植体细胞在Mebiol凝胶上培养。通过胸腺嘧啶核苷掺入研究分析培养细胞的增殖能力。对假定的角膜缘干细胞相关标志物和角膜分化标志物进行免疫染色，并通过逆转录（RT-PCR）进行确认。
角膜缘外植体在体外发生增殖。培养的细胞表达假定的角膜缘干细胞相关标志物（ABCG2和p63）、短暂扩增细胞标志物（连接蛋白43、整合素α9）和角膜分化标志物（K3）。RT-PCR证实了免疫组化数据。
我们的研究结果表明，合成聚合物Mebiol凝胶能够支持角膜缘外植体的增殖。培养的细胞表达假定的角膜缘干细胞相关标志物、短暂扩增细胞和角膜表型标志物。Mebiol凝胶可作为培养角膜缘外植体的支架。
