A 3D ex vivo mandible slice system for longitudinal culturing of transplanted dental pulp progenitor cells.
A 3D ex vivo mandible slice system for longitudinal culturing of transplanted dental pulp progenitor cells.
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DOI：
10.1002/cyto.a.22680
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发表时间：
2015-10
期刊：
影响因子：
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通讯作者：
Sloan AJ
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作者：
Colombo JS;Howard-Jones RA;Young FI;Waddington RJ;Errington RJ;Sloan AJ
利用间充质干细胞进行组织修复是再生医学的基础。然而，3D 组织基质如何使这些细胞保持静止状态，同时准备对组织损伤做出反应仍然相对未知。开发更多生理相关的 3D 模型将使我们能够更好地了解基质驱动因素以及对原位细胞谱系分化的影响。在这项研究中，我们开发了一种离体器官型大鼠下颌骨切片模型；一个技术定义的平台，用于培养和表征由 β-肌动蛋白启动子 (cGFP DPPC) 驱动的表达 GFP 的牙髓祖细胞。使用共聚焦显微镜，我们描述了原生环境如何影响移植到牙髓中的祖细胞。注射的 cGFP-DPPC 具有高度的活性，并且在下颌骨切片的独特区域中存在差异性增殖；在牙本质区域，cGFP-DPPC 显示出柱状形态，表明扩张和谱系分化。因此，我们证明了建立牙髓细胞微群落的系统能力，该微群落在牙齿中进行了表型修饰（“生物学”）；同时解决了技术挑战，使下颌骨切片能够在高内涵成像平台上进行访问（“多重”格式的生物学）。 © 2015 作者。由 Wiley 期刊公司出版
Harnessing mesenchymal stem cells for tissue repair underpins regenerative medicine. However, how the 3D tissue matrix maintains such cells in a quiescent state whilst at the same time primed to respond to tissue damage remains relatively unknown. Developing more physiologically relevant 3D models would allow us to better understand the matrix drivers and influence on cell‐lineage differentiation in situ. In this study, we have developed an ex vivo organotypic rat mandible slice model; a technically defined platform for the culture and characterization of dental pulp progenitor cells expressing GFP driven by the β‐actin promoter (cGFP DPPCs). Using confocal microscopy we have characterized how the native environment influences the progenitor cells transplanted into the dental pulp. Injected cGFP‐DPPCs were highly viable and furthermore differentially proliferated in unique regions of the mandible slice; in the dentine region, cGFP‐DPPCs showed a columnar morphology indicative of expansion and lineage differentiation. Hence, we demonstrated the systematic capacity for establishing a dental pulp cell‐micro‐community, phenotypically modified in the tooth (the “biology”); and at the same time addressed technical challenges enabling the mandible slice to be accessible on platforms for high‐content imaging (the biology in a “multiplex” format). © 2015 The Authors. Published by Wiley Periodicals, Inc.