Cell sheet technology exploits temperature responsive cell culture dishes (TRCDs) as versatile cell harvesting methods to yield contiguous cell monolayers robustly held together by cell–cell junctions, receptors, and endogenous extracellular matrix. More than 15 years of clinical data using autologous‐sourced cell sheets demonstrate enhanced therapeutic properties through increased cell retention at target tissue sites. Recently, several preclinical studies have also been reported using mesenchymal stem cell (MSC) sheets in wound healing, cardiac ischemia therapies, and pancreatic regeneration. However, optimized MSC sheet fabrication conditions have not yet been reported. In this study, we identified specific conditions for reliable human MSC sheet fabrication by comparing cell growth media supplements (fetal bovine serum [FBS] and human platelet lysate [hPL]). Human umbilical cord‐derived MSCs cultured in FBS and hPL exhibit different actin cytoskeletal structures related to their cell morphologies and adhesion. MSCs cultured in FBS media showed stable cell adhesion on TRCDs with flattened cell shapes and aligned actin cytoskeletal structure. This stable cell adhesion enables production of consistent MSC cell sheets, with controlled cell sheet detachment. Conversely, cell sheet fabrication in hPL media exhibits poor reproducibility being more sensitive to temperature‐ and culture time‐induced release due to weak cell adhesion. These findings suggest that stable MSC adhesion to TRCDs is important to reliable MSC sheet fabrication methods and that MSC growth media supplementation directly affects cell adhesion during culture.

中文翻译：

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稳定的细胞粘附影响间充质干细胞片的制造：胎牛血清和人血小板溶解产物的影响。

细胞片技术利用温度响应性细胞培养皿（TRCD）作为通用的细胞收集方法，以产生通过细胞-细胞连接，受体和内源性细胞外基质牢固结合在一起的连续细胞单层。使用自体来源细胞片的15年以上临床数据表明，通过增加靶组织部位的细胞滞留，可以增强治疗性能。最近，还报道了使用间充质干细胞（MSC）片进行伤口愈合，心脏缺血治疗和胰腺再生的一些临床前研究。但是，尚未报道优化的MSC片材制造条件。在这个研究中，我们通过比较细胞生长培养基补充剂（胎牛血清[FBS]和人血小板溶解产物[hPL]），确定了可靠的人MSC片材制造的特定条件。在FBS和hPL中培养的人脐带间充质干细胞表现出与其细胞形态和粘附有关的不同肌动蛋白细胞骨架结构。在FBS培养基中培养的MSC在TRCD上表现出稳定的细胞粘附性，细胞形状扁平且肌动蛋白细胞骨架结构对齐。这种稳定的细胞粘附力使得能够生产出一致的MSC细胞片，并具有受控的细胞片分离能力。相反，由于细胞粘附力弱，在hPL培养基中细胞片的制备显示出较差的重现性，对温度和培养时间诱导的释放更敏感。