Institut de Recherche en Hématologie et Transplantation (IRHT), Hôpital du Hasenrain, 87 Avenue d'Altkirch, 68100 Mulhouse, France.
CellProthera, 12, Rue du Parc, 68100 Mulhouse, France.
Int J Mol Sci. 2023 May 31;24(11):9551. doi: 10.3390/ijms24119551.
Ex vivo monitored human CD34 stem cells (SCs) injected into myocardium scar tissue have shown real benefits for the recovery of patients with myocardial infarctions. They have been used previously in clinical trials with hopeful results and are expected to be promising for cardiac regenerative medicine following severe acute myocardial infarctions. However, some debates on their potential efficacy in cardiac regenerative therapies remain to be clarified. To elucidate the levels of CD34 SC implication and contribution in cardiac regeneration, better identification of the main regulators, pathways, and genes involved in their potential cardiovascular differentiation and paracrine secretion needs to be determined. We first developed a protocol thought to commit human CD34 SCs purified from cord blood toward an early cardiovascular lineage. Then, by using a microarray-based approach, we followed their gene expression during differentiation. We compared the transcriptome of undifferentiated CD34 cells to those induced at two stages of differentiation (i.e., day three and day fourteen), with human cardiomyocyte progenitor cells (CMPCs), as well as cardiomyocytes as controls. Interestingly, in the treated cells, we observed an increase in the expressions of the main regulators usually present in cardiovascular cells. We identified cell surface markers of the cardiac mesoderm, such as kinase insert domain receptor (KDR) and the cardiogenic surface receptor Frizzled 4 (FZD4), induced in the differentiated cells in comparison to undifferentiated CD34 cells. The Wnt and TGF-β pathways appeared to be involved in this activation. This study underlined the real capacity of effectively stimulated CD34 SCs to express cardiac markers and, once induced, allowed the identification of markers that are known to be involved in vascular and early cardiogenesis, demonstrating their potential priming towards cardiovascular cells. These findings could complement their paracrine positive effects known in cell therapy for heart disease and may help improve the efficacy and safety of using ex vivo expanded CD34 SCs.
将体外监测的人 CD34 干细胞 (SCs) 注入心肌瘢痕组织中,已显示出对心肌梗死患者康复的实际益处。它们以前曾在临床试验中使用,结果令人充满希望,预计在严重急性心肌梗死后对心脏再生医学具有广阔前景。然而,关于它们在心脏再生治疗中的潜在疗效仍存在一些争议。为了阐明 CD34 SC 在心脏再生中的影响和贡献程度,需要更好地确定涉及它们潜在心血管分化和旁分泌分泌的主要调控因子、途径和基因。我们首先开发了一种方案,旨在促使从脐血中纯化的人 CD34 SC 向早期心血管谱系分化。然后,我们通过使用基于微阵列的方法,在分化过程中跟踪它们的基因表达。我们将未分化的 CD34 细胞与诱导分化的两个阶段(即第 3 天和第 14 天)的转录组进行了比较,同时还比较了人类心肌细胞祖细胞 (CMPCs) 和心肌细胞的转录组。有趣的是,在处理后的细胞中,我们观察到通常存在于心血管细胞中的主要调控因子的表达增加。我们鉴定了心脏中胚层的细胞表面标志物,例如激酶插入结构域受体 (KDR) 和心脏表面受体卷曲蛋白 4 (FZD4),这些标志物在分化细胞中被诱导表达,而在未分化的 CD34 细胞中则未被诱导表达。Wnt 和 TGF-β 途径似乎参与了这种激活。这项研究强调了有效刺激的 CD34 SC 表达心脏标志物的真实能力,并且一旦被诱导,就可以识别已知参与血管和早期心脏发生的标志物,证明它们对心血管细胞具有潜在的启动作用。这些发现可以补充它们在心脏病细胞治疗中已知的旁分泌积极作用,并可能有助于提高使用体外扩增的 CD34 SC 的疗效和安全性。