Tissue Harvesting Site and Culture Medium Affect Attachment, Growth, and Phenotype of Ex Vivo Expanded Oral Mucosal Epithelial Cells.
Tissue Harvesting Site and Culture Medium Affect Attachment, Growth, and Phenotype of Ex Vivo Expanded Oral Mucosal Epithelial Cells.
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DOI：
10.1038/s41598-017-00417-z
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发表时间：
2017-04-06
影响因子：
4.6
通讯作者：
Utheim TP
中科院分区：
文献类型：
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作者：
Islam R;Eidet JR;Badian RA;Lippestad M;Messelt E;Griffith M;Dartt DA;Utheim TP
培养的口腔黏膜上皮细胞移植是治疗角膜缘干细胞缺乏症的一种有效方法。为了改进培养方法，我们研究了四种培养基和组织收获部位对培养的Sprague-Dawley大鼠OMEC的外植体附着、生长和表型的影响。培养基或收获部位的选择均不影响外植体附着于培养孔的能力。Dulbecco改良Eagle培养基/Ham F12（DMEM）和Roswell Park Memorial Institute 1640培养基（RPMI）支持最大的细胞生长。培养6天后，LL外植体的生长倍数优于颊粘膜（BM）、HP和下唇过渡区（TZ）的外植体。在DMEM和RPMI中生长的培养物中检测到推定的干细胞标志物。在DMEM培养基中，TZ细胞的集落形成率高于LL、BM和HP。与RPMI相反，DMEM既表达假定的干细胞标记物Bmi-1，又产生细胞集落。我们的数据表明，OMEC从LL和TZ培养在DMEM中产生的未分化细胞具有高生长能力，因此是最有前途的角膜缘干细胞缺乏症的治疗。
Transplantation of cultured oral mucosal epithelial cells (OMECs) is a promising treatment strategy for limbal stem cell deficiency. In order to improve the culture method, we investigated the effects of four culture media and tissue harvesting sites on explant attachment, growth, and phenotype of OMECs cultured from Sprague-Dawley rats. Neither choice of media or harvesting site impacted the ability of the explants to attach to the culture well. Dulbecco’s modified Eagle’s medium/Ham’s F12 (DMEM) and Roswell Park Memorial Institute 1640 medium (RPMI) supported the largest cellular outgrowth. Fold outgrowth was superior from LL explants compared to explants from the buccal mucosa (BM), HP, and transition zone of the lower lip (TZ) after six-day culture. Putative stem cell markers were detected in cultures grown in DMEM and RPMI. In DMEM, cells from TZ showed higher colony-forming efficiency than LL, BM, and HP. In contrast to RPMI, DMEM both expressed the putative stem cell marker Bmi-1 and yielded cell colonies. Our data suggest that OMECs from LL and TZ cultured in DMEM give rise to undifferentiated cells with high growth capacity, and hence are the most promising for treatment of limbal stem cell deficiency.