Medical Center of Burn Plastic and Wound Repair, The First Affiliated Hospital of Nanchang University, Nanchang, China.
Department of Infectious Disease, The First Affiliated Hospital of Nanchang University, Nanchang, China.
Cell Transplant. 2024 Jan-Dec;33:9636897231218383. doi: 10.1177/09636897231218383.
Human embryonic stem cells (hESCs) are advantaged sources for large-scale and homogeneous mesenchymal stem/stromal cells (MSCs) generation. However, due to the limitations in high-efficiency procedures for hESC-MSCs induction, the systematic and detailed information of mesengenesis and early MSC development are largely obscure. In this study, we took advantage of the well-established twist-related protein 1 (TWIST1)-overexpressing hESCs and two small molecular cocktails (CHIR99021, decitabine) for high-efficient MSC induction. To assess the multidimensional biological and transcriptomic characteristics, we turned to cellular and molecular methods, such as flow cytometry (FCM), quantitative reverse transcription-polymerase chain reaction (qRT-PCR), tri-lineage differentiation, cytokine secretion analysis, transplantation for acute liver injury (ALI) management, and bioinformatics analyses (eg, gene ontology-biological processes [GO-BP], Kyoto Encyclopedia of Genes and Genomes [KEGG], HeatMap, and principal component analysis [PCA]). By combining TWIST1 overexpression (denoted as T) and the indicated small molecular cocktails (denoted as S), hESCs high-efficiently differentiated into MSCs (denoted as TS-MSCs, induced by T and S combination) within 2 weeks. TS-MSCs satisfied the criteria for MSC definition and revealed comparable tri-lineage differentiation potential and ameliorative efficacy upon ALI mice. According to RNA-sequencing (SEQ) analysis, we originally illuminated the gradual variations in gene expression pattern and the concomitant biofunctions of the programmed hESC-MSCs. Overall, our data indicated the feasibility of high-efficient generation of hESC-MSCs by TWIST1 and cocktail-based programming. The generated hESC-MSCs revealed multifaceted and biofunctions as adult BM-MSCs, which collectively suggested promising prospects in ALI management in future.
人类胚胎干细胞（hESCs）是大规模、均质间充质干细胞（MSCs）生成的有利来源。然而，由于高效 hESC-MSCs 诱导程序的限制，间充质发生和早期 MSC 发育的系统和详细信息在很大程度上仍不清楚。在这项研究中，我们利用已建立的 Twist 相关蛋白 1（TWIST1）过表达 hESC 和两种小分子鸡尾酒（CHIR99021、地西他滨）进行高效 MSC 诱导。为了评估多维生物学和转录组学特征，我们采用了细胞和分子方法，如流式细胞术（FCM）、实时定量逆转录聚合酶链反应（qRT-PCR）、三系分化、细胞因子分泌分析、急性肝损伤（ALI）管理的移植，以及生物信息学分析（例如，基因本体生物学过程 [GO-BP]、京都基因与基因组百科全书 [KEGG]、热图和主成分分析 [PCA]）。通过 TWIST1 过表达（表示为 T）和指定的小分子鸡尾酒（表示为 S）的组合，hESCs 在 2 周内高效地分化为 MSC（表示为 T 和 S 组合诱导的 TS-MSCs）。TS-MSCs 符合 MSC 定义的标准，并在 ALI 小鼠中显示出可比的三系分化潜力和改善功效。根据 RNA 测序（SEQ）分析，我们最初阐明了程序化 hESC-MSCs 基因表达模式的逐渐变化及其伴随的生物功能。总体而言，我们的数据表明，通过 TWIST1 和基于鸡尾酒的编程高效生成 hESC-MSCs 是可行的。生成的 hESC-MSCs 表现出多方面的和生物功能，作为成人 BM-MSCs，这共同表明在未来 ALI 管理中有广阔的前景。
