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A cost-effective method to prepare mesenchymal stem cells (MSCs) from human pluripotent stem cells

ABSTRACT: Induced mesenchymal stem cells (iMSCs) derived from human pluripotent stem cells (hPSCs) offer several advantages over tissue-derived mesenchymal stem cells (MSCs), including unlimited donor expansion, minimal batch-to-batch variation, a controllable pathogen microbiota, and the potential for large-scale production. However, producing iMSCs has several challenges, such as complex differentiation protocols, high costs, and poor reproducibility. Therefore, more effective differentiation methods are needed. In this study, iMSC differentiation was induced based on the classical method of using TGF-β signaling inhibitors. We discovered that due to the differences in the adhesiveness of gelatin to different cells, target cells can be rapidly selected and further differentiated to obtain high-purity iMSCs. This approach is more straightforward and less cost-effective than previous methods, producing iMSCs with higher purity and greater yield. We revealed that the DGEA active motif in gelatin produced effects similar to those of gelatin, playing an essential role in iMSC differentiation. In summary, our differentiation method addresses many of the issues associated with classical industrialization iMSC differentiation protocols and provides valuable insights into the production of iMSCs for clinical translation.

ORGANISM(S): Homo sapiens

PROVIDER: GSE289438 | GEO | 2026/05/01

REPOSITORIES: GEO

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Project description:Background: Radiation therapy is the standard of care for central nervous system tumours. Despite the success of radiation therapy in reducing tumour mass, irradiation (IR)-induced vasculopathies and neuroinflammation contribute to late-delayed complications, neurodegeneration, andpremature ageing in long-term cancer survivors. Mesenchymal stromal cells (MSCs) are adult stem cells that facilitate tissue integrity, homeostasis, and repair. Here, we investigated the potential of the iPSC-derived MSC(iMSC) secretome in immunomodulation and vasculature repair in response to radiation injury utilizing human cell lines. Methods: We generated iPSC-derived iMSC lines and evaluated the potential of their conditioned media (iMSC CM) to treat IR-induced injuries in human monocytes (THP1) and brain vascular endothelial cells (hCMEC/D3). We further assessed factors in the iMSC secretome, their modulation, and the molecular pathways they elicit. Results: Increasing doses of IR disturbed endothelial tube and spheroid formation in hCMEC/D3. When IR-injured hCMEC/D3 (IR ≤ 5 Gy) were treated with iMSC CM, endothelial cell viability, adherence, spheroid compactness, and proangiogenic sprout formation were significantly ameliorated, and IR-induced ROS levels were reduced. iMSC CM augmented tube formation in cocultures of hCMEC/D3 and iMSCs. Consistently, iMSC CM facilitated angiogenesis in a zebrafish model in vivo. Furthermore, iMSC CM suppressed IR-induced NFκB activation, TNF-α release, and ROS production in THP1 cells. Additionally, iMSC CM diminished NF-kB activation in THP1 cells cocultured with irradiated hCMEC/D3, iMSCs, or HMC3 microglial lines. The cytokine array revealed that iMSC CM contains the proangiogenic and immunosuppressive factors MCP1/CCL2, IL6, IL8/CXCL8, ANG (Angiogenin), GROα/CXCL1, and RANTES/CCL5. Common promoter regulatory elements were enriched in TF-binding motifs such as androgen receptor (ANDR) and GATA2. hCMEC/D3 phosphokinome profiling revealed increased expression of pro-survival factors, the PI3K/AKT/mTOR modulator PRAS40 and β-catenin in response to CM. The transcriptome analysis revealed increased expression of GATA2 in iMSCs and the enrichment of pathways involved in RNA metabolism, translation, mitochondrial respiration, DNA damage repair, and neurodevelopment. Conclusions: The iMSC secretome is a comodulated composite of proangiogenic and immunosuppressive factors that has the potential to alleviate radiation-induced vascular endothelial cell damage and immune activation.

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A cost-effective method to prepare mesenchymal stem cells (MSCs) from human pluripotent stem cells

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