Development of functional human oral mucosal epithelial stem/progenitor cell sheets using a feeder-free and serum-free culture system for ocular surface reconstruction.
Development of functional human oral mucosal epithelial stem/progenitor cell sheets using a feeder-free and serum-free culture system for ocular surface reconstruction.
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使用无馈物和无血清培养系统进行眼表面重建的功能性人口服粘膜上皮茎/祖细胞表的开发。
DOI：
10.1038/srep37173
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发表时间：
2016-11-14
影响因子：
4.6
通讯作者：
Kinoshita S
中科院分区：
文献类型：
--
作者：
Nakamura T;Yokoo S;Bentley AJ;Nagata M;Fullwood NJ;Inatomi T;Sotozono C;Yamagami S;Kinoshita S
利用组织工程化培养的口腔粘膜上皮细胞片（COMECS）进行眼表重建（OSR）是一种治疗严重眼表疾病的新方法。到目前为止，该技术使用外源性和未定义的组分，如小鼠来源的3 T3饲养细胞和胎牛血清。为了最大限度地减少人畜共患病感染或未知病原体传播的相关风险，从而为下一代治疗方式建立安全有效的方案，我们为COMECS方案开发了一种新技术，使用无饲养层和无血清（FFSF）培养系统。按照这种新的方案，COMECS表现出4-5层良好分层和分化的细胞，我们成功地产生了包括全克隆型干细胞的功能性COMECS。免疫组织化学证实FFSF COMECS中存在细胞连接（ZO 1，桥粒斑蛋白）、基底膜组装（胶原蛋白7，层粘连蛋白5）、分化（K13，K3）、增殖（Ki 67）和干/祖细胞（p75）的标志物。当移植到兔子的眼表面时，该组织存活长达2周。本研究是评估FFSF COMECS安全和理想OSR功能发展的第一步。
Ocular surface reconstruction (OSR) using tissue-engineered cultivated oral mucosal epithelial cell sheets (COMECS) is a promising newly developed treatment for patients with severe ocular surface disease. Until now, this technique has used exogenic and undefined components such as mouse-derived 3T3 feeder cells and fetal bovine serum. To minimize associated risks of zoonotic infection or transmission of unknown pathogens and so establish a safe and effective protocol for the next generation of treatment modality, we developed a novel technique for the COMECS protocol, using a feeder-free and serum-free (FFSF) culture system. Following this new protocol, COMECS exhibited 4–5 layers of well-stratified and differentiated cells, and we successfully produced functional COMECS that included holoclone-type stem cells. Immunohistochemistry confirmed the presence of markers for cell junction (ZO1, Desmoplakin), basement membrane assembly (Collagen 7, Laminin 5), differentiation (K13, K3), proliferation (Ki67) and stem/progenitor cells (p75) in the FFSF COMECS. When transplanted to the ocular surfaces of rabbits, the tissue survived for up to 2 weeks. This study represents a first step toward assessing the development of functional FFSF COMECS for safe and ideal OSR.
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影响因子：
13.7
作者：
Koizumi, N;Inatomi, T;Kinoshita, S
通讯作者：
Kinoshita, S
影响因子：
4.1
作者：
Nakamura, T;Inatomi, T;Kinoshita, S
通讯作者：
Kinoshita, S
影响因子：
158.5
作者：
Nishida, K;Yamato, M;Tano, Y
通讯作者：
Tano, Y
DOI：
10.1073/pnas.84.8.2302
发表时间：
1987-04-01
影响因子：
11.1
作者：
BARRANDON, Y;GREEN, H
通讯作者：
GREEN, H
影响因子：
13.7
作者：
Satake, Yoshiyuki;Higa, Kazunari;Shimazaki, Jun
通讯作者：
Shimazaki, Jun