通过团聚策略将人类干细胞诱导为成釉细胞。
Induction of human stem cells into ameloblasts by reaggregation strategy.
机构信息
Fujian Key Laboratory of Developmental and Neural Biology & Southern Center for Biomedical Research, College of Life Sciences, Fujian Normal University, Fuzhou, 350117, Fujian, P.R. China.
出版信息
Stem Cell Res Ther. 2024 Sep 27;15(1):332. doi: 10.1186/s13287-024-03948-1.
BACKGROUND
Human epithelium-derived stem cells and induced pluripotent stem cells (hiPSCs) possess the capability to support tooth formation and differentiate into functional enamel-secreting ameloblasts, making them promising epithelial-component substitutes for future human tooth regeneration. However, current tissue recombination approaches are not only technically challenging, requiring precise induction procedures and sophisticated microsurgery, but also exhibit low success rates in achieving tooth formation and ameloblastic differentiation.
METHODS
Suspended human keratinocyte stem cells (hKSCs) or cells from three hiPSC lines were directly mixed with dissociated embryonic mouse dental mesenchymal cells (mDMCs) that possess odontogenic potential in different proportions and reaggregated them to construct bioengineered tooth germs. The success rates of tooth formation and ameloblastic differentiation were confirmed after subrenal culture. The sorting capability, sequential development, and ameloblastic differentiation of stem cells were examined via GFP tracing, RT-PCR, and histological analysis, respectively.
RESULTS
Our reaggregation approach achieved an impressive success rate of more than 90% in tooth formation and 100% in ameloblastic differentiation when the chimeric tooth germs contained 1%~10% hKSCs or 5% hiPSCs. In addition, we observed that hiPSCs, upon exposure to mDMCs, initially transformed into epidermal cells, as indicated by KRT14 and CD29 expression, before progressing into dental epithelial cells, as indicated by SP6 and SHH expression. We also found that epithelial-derived hiPSCs, when reaggregated with mDMCs, were more favorable for tooth formation than their mesenchymal-derived counterparts.
CONCLUSIONS
This study establishes a simplified yet highly effective cell-cell reaggregation strategy for inducing stem cells to support tooth formation and differentiate into functional ameloblasts, paving the way for novel approaches for the development of stem cell-based tooth organoids and bioengineered tooth germs in vitro.
背景
人上皮细胞来源的干细胞和诱导多能干细胞（hiPSCs）具有支持牙齿形成并分化为功能性分泌釉质的成釉细胞的能力，使它们成为未来人类牙齿再生有前途的上皮成分替代物。然而，目前的组织重组方法不仅技术上具有挑战性，需要精确的诱导程序和复杂的显微手术，而且在实现牙齿形成和成釉细胞分化方面的成功率也很低。
方法
悬浮的人角质形成细胞干细胞（hKSCs）或来自三个 hiPSC 系的细胞与具有牙源性潜能的分离的胚胎小鼠牙间充质细胞（mDMCs）以不同比例直接混合并重新聚集以构建生物工程牙胚。在肾下培养后确认牙齿形成和成釉细胞分化的成功率。通过 GFP 追踪、RT-PCR 和组织学分析分别检查干细胞的分选能力、顺序发育和成釉细胞分化。
结果
我们的重新聚集方法在嵌合牙胚中包含 1%~10% hKSCs 或 5% hiPSCs 时，牙齿形成的成功率超过 90%，成釉细胞分化的成功率达到 100%。此外，我们观察到 hiPSCs 在暴露于 mDMCs 后，最初转化为表皮细胞，如 KRT14 和 CD29 的表达所示，然后转化为牙上皮细胞，如 SP6 和 SHH 的表达所示。我们还发现，与 mDMCs 重新聚集的上皮来源的 hiPSCs 比其间充质来源的 hiPSCs 更有利于牙齿形成。
结论
本研究建立了一种简化但非常有效的细胞-细胞重新聚集策略，用于诱导干细胞支持牙齿形成并分化为功能性成釉细胞，为基于干细胞的牙类器官和生物工程牙胚的体外开发开辟了新途径。
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