Mellott Adam J, Shinogle Heather E, Nelson-Brantley Jennifer G, Detamore Michael S, Staecker Hinrich
Department of Plastic Surgery, University of Kansas Medical Center, Kansas City, KS, 66160, USA.
Microscopy and Analytical Imaging Laboratory, University of Kansas, Lawrence, KS, 66045, USA.
Stem Cell Res Ther. 2017 Feb 28;8(1):41. doi: 10.1186/s13287-017-0505-6.
Use of decellularized tissues has become popular in tissue engineering applications as the natural extracellular matrix can provide necessary physical cues that help induce the restoration and development of functional tissues. In relation to cochlear tissue engineering, the question of whether decellularized cochlear tissue can act as a scaffold and support the incorporation of exogenous cells has not been addressed. Investigators have explored the composition of the cochlear extracellular matrix and developed multiple strategies for decellularizing a variety of different tissues; however, no one has investigated whether decellularized cochlear tissue can support implantation of exogenous cells.
As a proof-of-concept study, human Wharton's jelly cells were perfused into decellularized cochleae isolated from C57BL/6 mice to determine if human Wharton's jelly cells could implant into decellularized cochlear tissue. Decellularization was verified through scanning electron microscopy. Cocheae were stained with DAPI and immunostained with Myosin VIIa to identify cells. Perfused cochleae were imaged using confocal microscopy.
Features of the organ of Corti were clearly identified in the native cochleae when imaged with scanning electron microscopy and confocal microscopy. Acellular structures were identified in decellularized cochleae; however, no cellular structures or lipid membranes were present within the decellularized cochleae when imaged via scanning electron microscopy. Confocal microscopy revealed positive identification and adherence of cells in decellularized cochleae after perfusion with human Wharton's jelly cells. Some cells positively expressed Myosin VIIa after perfusion.
Human Wharton's jelly cells are capable of successfully implanting into decellularized cochlear extracellular matrix. The identification of Myosin VIIa expression in human Wharton's jelly cells after implantation into the decellularized cochlear extracellular matrix suggest that components of the cochlear extracellular matrix may be involved in differentiation.
脱细胞组织在组织工程应用中已变得很普遍，因为天然细胞外基质可提供必要的物理信号，有助于诱导功能性组织的修复和发育。在耳蜗组织工程方面，脱细胞耳蜗组织能否作为支架并支持外源细胞的植入这一问题尚未得到解决。研究人员已探索了耳蜗细胞外基质的组成，并开发了多种使各种不同组织脱细胞的策略；然而，没有人研究过脱细胞耳蜗组织是否能支持外源细胞的植入。
作为一项概念验证研究，将人脐带来源的间充质干细胞灌注到从C57BL/6小鼠分离的脱细胞耳蜗中，以确定人脐带来源的间充质干细胞是否能植入脱细胞耳蜗组织。通过扫描电子显微镜验证脱细胞效果。耳蜗用DAPI染色并用肌球蛋白VIIa进行免疫染色以鉴定细胞。灌注后的耳蜗用共聚焦显微镜成像。
用扫描电子显微镜和共聚焦显微镜成像时，在天然耳蜗中可清晰识别出柯蒂氏器的特征。在脱细胞耳蜗中鉴定出了无细胞结构；然而，通过扫描电子显微镜成像时，脱细胞耳蜗内不存在细胞结构或脂质膜。共聚焦显微镜显示，在用人脐带来源的间充质干细胞灌注后，脱细胞耳蜗中有细胞被阳性识别并附着。灌注后一些细胞阳性表达肌球蛋白VIIa。
人脐带来源的间充质干细胞能够成功植入脱细胞耳蜗细胞外基质。人脐带来源的间充质干细胞植入脱细胞耳蜗细胞外基质后肌球蛋白VIIa表达的鉴定表明，耳蜗细胞外基质的成分可能参与分化。
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