Talebi Mehdi, Nozad Charoudeh Hojjatollah, Movassaghpour Akbari Ali Akbar, Baradaran Behzad, Kazemi Tohid
Department of Applied Cell Sciences, School of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran.
Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Adv Pharm Bull. 2020 Sep;10(4):617-622. doi: 10.34172/apb.2020.074. Epub 2020 Aug 9.
Because of different potentials of T-cell subtypes in T-cell based cellular immunotherapy approaches such as CAR-T cell therapies; Regarding the high cost of the serum-free specific culture media, having distinct control on T-cell subset activation, expansion and differentiation seem crucial in T-cell expansion step of cell preparation methods. By the way, there was no clear data about the effect of acellular Wharton's Jelly (AWJ) on T-cells expansion, activation or differentiation status. So, we have launched to study the effect of AWJ on T-cell's immunobiological properties. CD3+ T-cells were isolated from healthy bone marrow allogeneic donors, sorted by FACS method and cultured on either routine phyto-hemagglutinin complemented and different concentrations of AWJ, lag phase and doubling time of the cells calculated from cell growth curve. After 3, 7 and 14-days T-cell subtypes cell markers and cell activity related genes expression rate have been evaluated by flow cytometry and real-time polymerase chain reaction (PCR) methods respectively. AWJ in a 1:1 ratio compared with contemporary lymphocyte culture media showed significant activating and proliferative capacities. The introduced condition has not affected the frequency of CD4+ subpopulation of T-cells, but significantly increased even CD8+ cells and immune-activator genes in T-cells. The regulatory and memory subsets of T-cells in this study have not affected significantly. the study results revealed that AWJ can be utilized as a supportive substance to increase the memory properties of the T-cells, gives control to design a selective medium for expanding and differentiating memory T-cells, relatively.
由于在基于T细胞的细胞免疫疗法(如嵌合抗原受体T细胞疗法)中T细胞亚型具有不同的潜能;考虑到无血清特异性培养基成本高昂,在细胞制备方法的T细胞扩增步骤中,对T细胞亚群的激活、扩增和分化进行明确控制显得至关重要。顺便说一句,关于脱细胞沃顿胶(AWJ)对T细胞扩增、激活或分化状态的影响,尚无明确数据。因此,我们着手研究AWJ对T细胞免疫生物学特性的影响。从健康的异基因骨髓供体中分离出CD3+ T细胞,通过荧光激活细胞分选(FACS)方法进行分选,并在常规植物血凝素补充剂和不同浓度的AWJ上培养,根据细胞生长曲线计算细胞的延迟期和倍增时间。分别在3天、7天和14天后,通过流式细胞术和实时聚合酶链反应(PCR)方法评估T细胞亚型细胞标志物和细胞活性相关基因的表达率。与当代淋巴细胞培养基相比,1:1比例的AWJ显示出显著的激活和增殖能力。所引入的条件未影响T细胞CD4+亚群的频率,但显著增加了T细胞中CD8+细胞甚至免疫激活基因的数量。本研究中T细胞的调节性和记忆性亚群未受到显著影响。研究结果表明,AWJ可作为一种支持性物质来增强T细胞的记忆特性,相对而言,有助于设计一种用于扩增和分化记忆T细胞的选择性培养基。