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INTRODUCTION

It is important to prepare 'hypoimmunogenic' or 'universal' human pluripotent stem cells (hPSCs) with gene-editing technology by knocking out or in immune-related genes, because only a few hypoimmunogenic or universal hPSC lines would be sufficient to store for their off-the-shelf use. However, these hypoimmunogenic or universal hPSCs prepared previously were all genetically edited, which makes laborious processes to check and evaluate no abnormal gene editing of hPSCs.

METHODS

Universal human-induced pluripotent stem cells (hiPSCs) were generated without gene editing, which were reprogrammed from foetal stem cells (human amniotic fluid stem cells) with mixing 2-5 allogenic donors but not with single donor. We evaluated human leucocyte antigen (HLA)-expressing class Ia and class II of our hiPSCs and their differentiated cells into embryoid bodies, cardiomyocytes and mesenchymal stem cells. We further evaluated immunogenic response of transient universal hiPSCs with allogenic mononuclear cells from survival rate and cytokine production, which were generated by the cells due to immunogenic reactions.

RESULTS

Our universal hiPSCs during passages 10-25 did not have immunogenic reaction from allogenic mononuclear cells even after differentiation into cardiomyocytes, embryoid bodies and mesenchymal stem cells. Furthermore, the cells including the differentiated cells did not express HLA class Ia and class II. Cardiomyocytes differentiated from transient universal hiPSCs at passage 21-22 survived and continued beating even after treatment with allogenic mononuclear cells.

INTRODUCTION

METHODS

RESULTS

简介

通过敲除或编辑免疫相关基因，用基因编辑技术制备“低免疫原性”或“通用”人多能干细胞（hPSC）非常重要，因为只需少数几个低免疫原性或通用 hPSC 系就足以储存以备随时使用。然而，之前制备的这些低免疫原性或通用 hPSC 都是经过基因编辑的，这使得检查和评估 hPSC 中没有异常基因编辑的过程变得繁琐。

方法

无需基因编辑即可生成通用的人诱导多能干细胞（hiPSC），这些细胞是由混合了 2-5 个同种异体供体的胎儿干细胞（人羊水干细胞）重新编程而来，而非单一供体。我们评估了我们的 hiPSC 及其分化为胚状体、心肌细胞和间充质干细胞的细胞表达人类白细胞抗原（HLA）的 I 类和 II 类。我们进一步评估了瞬时通用 hiPSC 与异体单核细胞之间的免疫反应，通过异体单核细胞的存活率和细胞因子产生来评估，这是由于免疫反应导致细胞产生的。