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

Here, we introduce a novel chemically induced dedifferentiation approach that reverses the cellular development from a differentiated state to progenitor state with reduced senescence features, 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. we use small-molecule combiantions to induce the dedifferentiation of human somatic cells to progenitors and then characterized the features of the progenitors.

本研究提出一种全新的化学诱导去分化（chemically induced dedifferentiation）方法，可将细胞从分化状态逆转至衰老特征减弱的祖细胞状态，并实现从成人源间充质基质细胞（human adult mesenchymal stromal cells）与真皮成纤维细胞（dermal fibroblasts）诱导生成肢芽祖细胞（limb bud progenitors）。此类化学诱导肢芽细胞（chemically induced limb bud cells，简称hCiLB细胞）与人类胚胎肢芽细胞具有极高的转录组相似性。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，相较于亲本细胞，hCiLB细胞具备更强的成骨成软骨分化能力，可为构建功能性关节软骨提供全新研究平台。本研究结果同时揭示了一种潜在治疗策略：通过将成熟细胞逆转至可扩增祖细胞的发育通路，实现受损人体组织的修复。本研究采用小分子组合诱导人体体细胞向祖细胞去分化，并对所得祖细胞的特征进行了系统表征。