Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, 81744-176, Iran.
Division of Laboratory Hematology and Blood Banking, Department of Medical Laboratory Sciences, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran.
Cell Tissue Bank. 2023 Jun;24(2):485-494. doi: 10.1007/s10561-022-10048-y. Epub 2022 Nov 25.
Hematopoietic stem cells (HSCs) can be isolated through umbilical cord blood (UCB), which can be used for HSC transplantation. Despite many advantages, the low number of UCB CD34 cells lead to delayed engraftment. Ex-vivo CD34HSC expansion is a potentially safe approach to increasing CD34 cell numbers. The NLR family of pyrin domain-containing 3 (NLRP3) is an intracellular protein that plays an essential role in the innate immune response. Several blood cell types, HSCs and progenitor cells (HSPCs) express the NLRP3 inflammasome complex genes and participate in the development and proliferation of HSPCs. In this study, magnetic-activated cell sorting (MACS) beads isolated CD34HSCs. The cell purity was evaluated by flow cytometry. CD34 cells, under the influence of different doses of glucose, MCC950 were cultured for seven days. The qRT-PCR was used to evaluate gene expression. The results showed that in the culture medium treated with glucose concentrations, the expression of the NLRP3 inflammasome complex genes and the amount of CD34 cells increased by more than 50%. In contrast, genes expression and the number of CD34 cells in the culture medium treated with MCC950 decreased. UCB is a source of new therapeutic methods. This study demonstrates the relationship between glucose and the activation of the NLRP3 inflammasome. Based on these results, glucose causes the expansion of CD34HSCs through its effect on HSCs in simultaneous culture.
造血干细胞(HSCs)可通过脐带血(UCB)分离,可用于 HSC 移植。尽管有许多优点,但 UCB CD34 细胞数量低导致植入延迟。体外 CD34+HSC 扩增是增加 CD34 细胞数量的一种潜在安全方法。NLR 家族包含 pyrin 结构域的 3(NLRP3)是一种在先天免疫反应中起重要作用的细胞内蛋白。几种血细胞类型、HSCs 和祖细胞(HSPCs)表达 NLRP3 炎性体复合物基因,并参与 HSPCs 的发育和增殖。在这项研究中,通过磁激活细胞分选(MACS)珠分离 CD34+HSCs。通过流式细胞术评估细胞纯度。在不同剂量葡萄糖、MCC950 的影响下培养 CD34 细胞七天。采用 qRT-PCR 评估基因表达。结果表明,在葡萄糖浓度处理的培养基中,NLRP3 炎性体复合物基因的表达和 CD34 细胞数量增加了 50%以上。相比之下,MCC950 处理的培养基中基因表达和 CD34 细胞数量减少。UCB 是新治疗方法的来源。本研究表明葡萄糖与 NLRP3 炎性体的激活之间存在关系。基于这些结果,葡萄糖通过对同时培养的 HSCs 的作用导致 CD34+HSCs 的扩增。