Naseri Mobaraki Sepideh, Abroun Saeid, Atashi Amir, Kaviani Saeid
Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Stem Cell and Tissue Engineering Research Center, Shahroud University of Medical Sciences, Shahroud, Iran.
Cell J. 2023 Mar 7;25(3):184-193. doi: 10.22074/cellj.2022.557607.1084.
Umbilical cord blood (UCB) is an accessible and effective alternative source for hematopoietic stem cell (HSC) transplantation. Although the clinical application of UCB transplantation has been increased recently, quantitative limitation of HSCs within a single cord blood unit still remains a major hurdle for UCB transplantation. In this study we used microcarrier beads to evaluate the expansion of UCB-derived HSCs in co-cultured with UCB-derived mesenchymal stem cells (MSC).
In this experimental study, we used microcarrier beads to expand UCB-derived MSCs. We investigated the simultaneous co-culture of UCB-derived CD34 cells and MSCs with microcarrier beads to expand CD34 cells. The colony forming capacity and stemness-related gene expression on the expanded CD34 cells were assessed to determine the multipotency and self-renewal of expanded cells.
Our results indicated that the microcarrier-based culture significantly increased the total number and viability of UCB-derived MSCs in comparison with the monolayer cultures during seven days. There was a significant increase in the UCB-derived CD34+ cells expanded in the presence of microcarrier beads in this co-culture system. The expanded UCB-derived CD34+ cells had improved clonogenic capacity, as evidenced by higher numbers of total colony counts, granulocyte, erythrocyte, monocyte, megakaryocyte colony forming units (CFU-GEMM), and granulocyte-monocyte colony forming units (CFU-GM). There were significantly increased expression levels of key regulatory genes (CXCR4, HOXB4, BMI1) during CD34 cells self-renewal and quiescence in the microcarrier-based co-culture.
Our results showed that the increase in the expansion and multipotency of CD34 cells in the microcarrierbased co-culture can be attributed to the enhanced hematopoietic support of UCB-derived MSCs and improved cell-cell interactions. It seems that this co-culture system could have the potential to expand primitive CD34 cells.
脐带血（UCB）是造血干细胞（HSC）移植的一种可获取且有效的替代来源。尽管近年来UCB移植的临床应用有所增加，但单个脐带血单位内HSC数量的限制仍是UCB移植的主要障碍。在本研究中，我们使用微载体珠评估与UCB来源的间充质干细胞（MSC）共培养时UCB来源的HSC的扩增情况。
在本实验研究中，我们使用微载体珠扩增UCB来源的MSC。我们研究了将UCB来源的CD34细胞和MSC与微载体珠同时共培养以扩增CD34细胞。评估扩增后的CD34细胞的集落形成能力和干性相关基因表达，以确定扩增细胞的多能性和自我更新能力。
我们的结果表明，与单层培养相比，基于微载体的培养在7天内显著增加了UCB来源的MSC的总数和活力。在该共培养系统中，存在微载体珠时，UCB来源的CD34 +细胞有显著增加。扩增后的UCB来源的CD34 +细胞具有更高的克隆形成能力，总集落计数、粒细胞、红细胞、单核细胞、巨核细胞集落形成单位（CFU - GEMM）和粒细胞 - 单核细胞集落形成单位（CFU - GM）数量增加证明了这一点。在基于微载体的共培养中，CD34细胞自我更新和静止期间关键调控基因（CXCR4、HOXB4、BMI1）的表达水平显著增加。
我们的结果表明，基于微载体的共培养中CD34细胞扩增和多能性的增加可归因于UCB来源的MSC增强的造血支持和改善的细胞间相互作用。似乎这种共培养系统有可能扩增原始CD34细胞。
