Microvesicles from Human Immortalized Cell Lines of Endothelial Progenitor Cells and Mesenchymal Stem/Stromal Cells of Adipose Tissue Origin as Carriers of Bioactive Factors Facilitating Angiogenesis

Abstract

Endothelial progenitor cells (EPCs) and mesenchymal stem/stromal cells (MSCs) are associated with maintaining tissue homeostasis and tissue repair. Both types of cells contribute to tissue regeneration through the secretion of trophic factors (alone or in the form of microvesicles). This study investigated the isolation and biological properties of microvesicles (MVs) derived from human immortalized MSC line HATMSC1 of adipose tissue origin and EPC line. The human immortalized cell line derived from the adipose tissue of a patient with venous stasis was established in our laboratory using the hTERT and pSV402 plasmids. The human EPC line originating from cord blood (HEPC-CB.1) was established in our previous studies. Microvesicles were isolated through a sequence of centrifugations. Analysis of the protein content of both populations of microvesicles, using the Membrane-Based Antibody Array and Milliplex ELISA showed that isolated microvesicles transported growth factors and pro- and antiangiogenic factors. Analysis of the miRNA content of isolated microvesicles revealed the presence of proangiogenic miRNA (miR-126, miR-296, miR-378, and miR-210) and low expression of antiangiogenic miRNA (miR-221, miR-222, and miR-92a) using real-time RT-PCR with the TaqMan technique. The isolated microvesicles were assessed for their effect on the proliferation and proangiogenic properties of cells involved in tissue repair. It was shown that both HEPC-CB.1- and HATMSC1-derived microvesicles increased the proliferation of human endothelial cells of dermal origin and that this effect was dose-dependent. In contrast, microvesicles had a limited impact on the proliferation of fibroblasts and keratinocytes. Both types of microvesicles improved the proangiogenic properties of human dermal endothelial cells, and this effect was also dose-dependent, as shown in the Matrigel assay. These results confirm the hypothesis that microvesicles of HEPC-CB.1 and HATMSC1 origin carry proteins and miRNAs that support and facilitate angiogenic processes that are important for cutaneous tissue regeneration.

Suppression of Ovarian Cancer Cell Growth by AT-MSC Microvesicles

“…MVs were isolated according to the well-established protocol developed in our laboratory [45] based on the procedure introduced in the previous study [46]. HATMSC2 cells were cultured in multi-layer cell culture flasks (Nunc TripleFlasks, Thermo Scientific, Carlsbad, CA, USA) using DMEM + 10% FBS until they reached 75% confluence.…”

“…ES-2 and OAW-42 cells were seeded into a Lab-Tek II Chambered # 1.5 Coverglass system (Nalge Nunc International, Naperville, IL, USA) at a density of 15 × 10 3 cells per chamber. Fluorescence staining of the MVs was performed, as established in our recent study [45]. After washing with PBS, the MVs were resuspended in the DMEM + 10% FBS and OptiMEM GlutaMax + 3% FBS media (mixed in equal proportions), and added to the cells at a ratio of 5:1 (5 MVs per cell) and 10:1 (10 MVs per cell).…”

Perspectives on mesenchymal stem/progenitor cells and their derivates as potential therapies for lung damage caused by COVID-19

“…The trophic properties of MSCs are associated with the secretion of growth factors and chemokines, such as TGF-α, TGF-β, hepatocyte growth factor (HGF), epithelial growth factor (EGF), insulin-like growth factor 1, bFGF, vascular endothelial growth factor (VEGF), angiopoietin-1 (Ang-1), and other bioactive factors involved in cell proliferation and angiogenesis, as confirmed by many studies[ 31 , 36 ] including research conducted by the author of this article[ 33 , 37 , 38 ].…”

Identification of key pathways and hub genes in the myogenic differentiation of pluripotent stem cell: a bioinformatics and experimental study

“…[5][6][7]. Stem cells are promising cells that have the potency of multidirectional differentiation and proliferation and are widely expected to be used in the field of tissue repair and regeneration [8,9]. In the muscle regeneration field, stem cells also showed vigorous potency [10].…”