High glucose condition limited the angiogenic/cardiogenic capacity of murine cardiac progenitor cells in in vitro and in vivo milieu

Abstract

Murine c-kit cardiac cells were isolated and enriched by magnetic activated cell sorting technique. c-kit cells viability and colony-forming activity were evaluated by MTT and clonogenic assay. c-kit cells were exposed to endothelial, pericyte, and cardiomyocyte induction media containing 30mM glucose for 7 days. We monitored the level of endothelial (VE-cadherin, CD31, and vWF), pericyte (NG , α-SMA, and PDGFR-β), and cardiomyocyte markers (cTnT) using flow cytometry, real-time Polymerase Chain Reaction (PCR), and Enzyme-Linked Immunosorbent Assay (ELISA) analyses. Ultrastructural changes were studied by transmission electron microscopy (TEM) in cells treated with 5-Azacytidine and 30mM glucose. Matrigel plug assay was performed to determine the angio/cardiogenic property of c-kit cells in a diabetic mouse model. Glucose of 30mM decreased c-kit cells viability and clonogenicity (P < 0.05). The transdifferentiation capacity of c-kit cells into the endothelial lineage, pericytes, and cardiomyocytes were reduced through the inhibition of related genes (P < 0.05). TEM analysis revealed cardiomyocyte differentiation rate in c-kit cells coincided with an increased intracellular lipid accumulation and reduced number of mitochondria. Similar to in vitro condition, the angiogenic capacity of c-kit cells was aborted in vivo indicated by reduced NG , α-SMA, CD31, and vWF levels. High glucose condition reduces the angio/cardiogenic capacity of cardiac c-kit cells in vitro and in vivo. SIGNIFICANCE OF THE STUDY: High glucose condition seen in diabetes mellitus could affect the regenerative potential of cardiac tissue. The current experiment showed that the exposure of murine cardiac progenitor cells (CD117 cells) to condition containing 30mM glucose could decrease the differentiation properties into endothelial cells, pericytes, and mature cardiomyocytes in vitro and in vivo. Our finding confirmed that the angiogenic/cardiogenic potential cardiac progenitor cells decrease under treatment with high glucose content as seen in the diabetic condition.

C‐Kit+ progenitors restore rat asthmatic lung function by modulation of T‐bet and GATA‐3 expression

“…130‐091‐224; Miltenyi Biotec,Bergisch Gladbach, Germany) according to the manufacturer's recommendations. Then the cells were passed through LS columns (Miltenyi Biotec, Bergisch Gladbach, Germany), and the Kit + and Kit − cells were collected separately and subjected to different analyses (Khaksar et al., 2018).…”

“…To confirm the phenotype of enriched cells, we performed flow cytometry to measure the percentage of Kit + cells after the MACS procedure (Khaksar et al., 2018). In brief, cells from the negative and positive groups were incubated with mouse‐anti‐human CD117 (Kit + ) at 4°C for 30 min.…”

Intratracheal administration of bone marrow c-kit+ cells offered hopes in ameliorating asthmatic pathologies via the control of miRNA-133 and -126

“…Then, cells were incubated with mouse-anti human c-Kit microbead (Miltenyi Biotech, Germany) according to manufacturer's instructions. By passing cells through the LS columns (Miltenyi Biotec, Germany) and c-Kit + and c-Kitcells were isolated and used for different analyses [18].…”

“…The multipotentiality of isolated cells was studied after MACS by ow cytometry analysis [18].In short, cells both groups were incubated with FITC-conjugated mouse-anti human CD117 (c-kit + ) at 4˚C for 30 min. The samples were analyzed by BD FACS Calibur and raw data processed using FlowJo software (Ver.…”

Distinct role of autophagy on angiogenesis: highlights on the effect of autophagy in endothelial lineage and progenitor cells

“…Angiogenesis is a crucial phenomenon for maintaining blood nourishment under the normal condition that also participates in the regeneration of ischemic cardiac tissue or impaired peripheral vascular [ 114 , 115 ]. In contrast, the tumor progression and metastasis could be enhanced by promoting angiogenesis under pathological conditions [ 116 ].…”