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A Study of novel mechanism of hypoxic preconditioned CD34+cells improving therapeutic efficacy for critical limb ischemia: role of exosomal miRNA-378a-5p

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Background & Aim CD34+cells transplantation has shown therapeutic angiogenesis efficacy in treating critical limb ischemia. Hypoxic preconditioning can increase the proangiogenic ability. As the main component of the paracrine secretion… Click to show full abstract

Background & Aim CD34+cells transplantation has shown therapeutic angiogenesis efficacy in treating critical limb ischemia. Hypoxic preconditioning can increase the proangiogenic ability. As the main component of the paracrine secretion of CD34+ cells, CD34+ cell exosome (CD34-Exo) plays an important role. In this research, we aim to investigate the proangiogenic mechanism of hypoxic preconditioned CD34 exosome (HCC34-Exo) at the level of exsosomal miRNA. Methods, Results & Conclusion RNAseq screen showed 3.40±2.85 (means±standrad deviations) folds (n=6) of mi-RNA-387a-5p in the HCC34-Exo than in the normoxic preconditioned CD34+ cell exosomes (NCC34-Exo), and this was validated by qRT-PCR. In vitro experiments such as transwell co-culture model, exosome secretion-blocked experiment, exosome tracking experiment demonstrated that miRNA-378a-5p was directly transferred from the CD34+ cells through exosomes of CD34+ cells to human umbilical vein endothelial cells (HUVECs). Tube formation experiment demonstrated the enhancement of the HUVECs function, significant increase was observed concerning branching points in HCC34-Exo group, which equals to 2.15±0.46 folds of that in blank control group, 1.80±0.62 folds of that in anti-miRNA-378 HCC34-Exo group and 1.32±0.34 folds of that in NCC-Exo group (all p Our results have showed that HCC34 might promote therapeutic angiogenesis in ischemic tissue through exosomal miR-378-5p. Identification of the mechanism will provide important theoretical evidence for the therapeutic efficacy of HCC34 and its exsosomal miRNA in treating critical limb ischemia and may offer a new idea for potential treatment strategies in the future.

Keywords: cd34 cells; efficacy; limb ischemia; preconditioned cd34; critical limb

Journal Title: Cytotherapy
Year Published: 2020

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