Abstract
Purpose: 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. Methods: 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. Results: 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. Conclusion: 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.
Highlights
Cellular products process engineering needs having exact knowledge about every step of the known process and having control over each of them
Because of different potentials of T-cell subtypes in T-cell based cellular immunotherapy approaches such as chimeric antigen receptors (CARs)-T cell therapies; Regarding the high cost of the serumfree 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
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 acellular Wharton’s Jelly (AWJ), lag phase and doubling time of the cells calculated from cell growth curve
Summary
Cellular products process engineering needs having exact knowledge about every step of the known process and having control over each of them. As well, based on world major GMP (Good Manufacturing Practice) regulatory the more minimizing the manipulating cellular products the secure the products. One of the cellular products branches immune-cell therapy, and especially T-cell based adaptive immune therapy approaches have growing interests during recent years. The CAR-T cell therapy approaches are one of the most interesting ones that utilize the antibody-based chimeric antigen receptors (CARs) on T-cells to recognition tumor-specific antigens for effectively removing hematologic and solid tumor cells.[1,2] CAR-T cell (primarily called T body) was firstly described at Weizmann Institute of Science in Israel by Eshhar et al.[3] T-cells naturally are composing of different subtypes that will be described below and the composition of the in an individual at different infective/inflammations and/or reactive states results in various immunological, even activating and suppressive conditions. Producing the CAR-T cells will utilize different subtypes of T-cells even effector or memory cells, the importance of this approach highlighted in Louis et al and Xu et al reports.[4,5] For example, Gattinoni and colleagues reported that CAR-T-cells produced from T memory Stem cell (TSCM) that highly expressing CCR-7, CD95 and CD62L, shown a more durable and effective anti-tumoral effect in comparison with central memory T cells.[6,7] Generally, CD4+ T-cell subtypes, Th1, Th2, Th9, Th17, Th22, Treg
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