中国输血杂志2025，Vol.38Issue(5)：644-651,8.DOI:10.13303/j.cjbt.issn.1004-549x.2025.05.008
造血类器官来源NK支持细胞HO-hMSCs的获取与功能研究
Isolation andfunctional characterization of HO-hMSCs as NK-supportive cells derived from hematopoietic organoids
摘要
Abstract
Objective In in vitro systems for differentiating and expanding natural killer(NK)cells,feeder cells pro-vide essential cell-cell contact and paracrine signals that drive precursor proliferation and terminal maturation.However,ex-isting xenogeneic feeder cells or tumor-derived genetically modified feeder cells pose risks of residual immunogenicity and malignant transformation,limiting clinical use.This study aims to develop a humanized mesenchymal-like stromal cell(hematopoietic organoid-derived human mesenchymal stromal cells,HO-hMSCs)derived from iPSC-based hematopoietic or-ganoids,and elucidate its mechanisms of NK-supportive activity to enable a safe,efficient platform for clinical-grade NK cell production.Methods Human induced pluripotent stem cells(iPSCs)were differentiated into hematopoietic or-ganoids,from which HO-hMSCs were isolated.Flow-cytometric phenotyping and bulk RNA-sequencing were performed to compare HO-hMSCs with umbilical cord-derived MSCs(UC-hMSCs).The effect of HO-hMSCs on NK cell differentiation efficiency(CD3-CD56+)and effector maturation(CD16 expression)were assessed by co-culture experiments,using UC-hMSCs as control.Results 1)Hematopoietic organoid induction and NK differentiation:iPSCs were induced to form hema-topoietic organoids using cytokine cocktails,which further differentiated into high-purity CD45+CD56+NK cells[(82.8%±12.07)%efficiency on day 21].2)HO-hMSC characteristics:HO-hMSCs exhibited upregulated expression of Notch path-way ligands(DLL4,JAG1,4.06-8.04-fold),homeobox genes(HOXA3,HOXA5,log2 FC=1.28 and 1.44),and key regulators of NK development(GATA3,BCL11A)and cytokine receptors(IL7R,IL27RA,6.76 to 13.34-fold increase).3)Functional validation:Compared to UC-hMSCs,HO-hMSCs co-culture significantly enhanced NK cell proportion by 30.5%(P<0.05)and increased CD16 positivity(+20.5%).Conclusion This study for the first time reveals that human hematopoietic organoid-derived HO-hMSCs possess potent hematopoietic niche-supportive activity.It provides a humanized,feeder-free platform for robust clinical-grade NK cell production and expands the translational utility of organoid technologies in cell therapy.关键词
造血类器官/间充质细胞/饲养层细胞/NK分化Key words
hematopoietic organoid/mesenchymal stromal cells/feeder layer cells/NK differentiation分类
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唐诗丽,林碧萱,黄恩霞,赫英,薛原,张勇刚..造血类器官来源NK支持细胞HO-hMSCs的获取与功能研究[J].中国输血杂志,2025,38(5):644-651,8.基金项目
四川省科技厅省级科技计划项目(2024NSFSC0618) （2024NSFSC0618）
