Generation of Human Expandable Limb Bud Cells via Chemically Induced Dedifferentiation Strategy [bulk RNA-seq]. Generation of Human Expandable Limb Bud Cells via Chemically Induced Dedifferentiation Strategy [bulk RNA-seq]

Here, we introduce a novel chemically induced dedifferentiation approach that reverses the cellular development from a differentiated state to progenitor state, conferring the generation of limb bud progenitors from human adult mesenchymal stromal cells and dermal fibroblasts. These chemically induced limb bud cells (hCiLB cells) show a high degree of transcriptomic similarity to human embryonic limb bud cells. We found that hCiLB cells can undergo extensive expansion, while maintaining population homogeneity and long-term self-renewal capacity. Moreover, hCiLB cells exhibit increased osteochondrogenic differentiation ability than their parental cells, providing a new platform for the development of authentic articular cartilage. These results also highlight a potential therapeutic approach for the repair of damaged human tissues through reversal of developmental pathways from mature cells to expandable progenitor cells. Overall design: we use small-molecule combiantions to induce the dedifferentiation of human somatic cells to progenitors and then characterized the features of the progenitors.

本研究报道一种全新的化学诱导去分化方法，该方法可逆转细胞发育进程，将已分化细胞重编程为祖细胞状态，并可从人类成人间充质基质细胞（mesenchymal stromal cells）与真皮成纤维细胞（dermal fibroblasts）中诱导生成肢芽祖细胞（limb bud progenitors）。这类化学诱导肢芽细胞（chemically induced limb bud cells, hCiLB cells）的转录组特征与人类胚胎肢芽细胞高度相似。研究发现，hCiLB细胞可实现大规模扩增，同时维持群体均一性与长期自我更新能力。此外，hCiLB细胞相较于其亲本细胞，成骨软骨分化能力显著提升，为构建功能性原生关节软骨提供了全新研究平台。本研究结果同时揭示了一种潜在治疗策略：通过逆转成熟细胞向可扩增祖细胞的发育通路，实现受损人体组织的修复。实验整体设计：采用小分子组合诱导人类体细胞向祖细胞去分化，随后对该祖细胞的各项特征进行表征分析。