Investigating the function of combining induced rat monocytes-derived bone marrow-haemopoietic stem cell (rat BM-HSCs) with LPS and rat bone marrow-mesenchymal stem cell (rat BM-MSCs) was to analyze the acceleration o...Investigating the function of combining induced rat monocytes-derived bone marrow-haemopoietic stem cell (rat BM-HSCs) with LPS and rat bone marrow-mesenchymal stem cell (rat BM-MSCs) was to analyze the acceleration of homing process mechanism in injured pancreas. Mononucleated stem cells were isolated from aspirated whole rat BM using ficoll and cultured in α-MEM complete growth medium in 10 cm petridish. After two days, adherent cells after washing twice in petridish were added α-MEM growth medium and then mesenchymal cells were characterized using CD105 marker in third passage and labeled PKH26. Then haemopoietic stem cells (HSCs) were isolated with magnetic beads CD34+ and differentiated in vitro, and then induced monocytes with LPS. Animal experiment used 28 male Wistar rats, and divided them into 4 groups. After transplantation combined, both cells between monocyte derived HSc (mHSCs) and rat BM-MSC were analyzed expression of pair box gen 4 (Pax4), pancreatic and duodenal homeobox (Pdx1), C-peptide using immunohistochemistry, then secretion of insulin and C-peptide analyzed using indirect ELISA. Results showed that the expressions of Pax4, Pdx1, C-peptide found in the surface membrane cell of pancreatic cell, and secreted C-peptide and insulin were shown significant (P < 0.05) in transplanted group 2, 3 and 4, but in group 3 were transplanted with combined cells more dominant than non-combined cells. Conclusions suggested that combining of induced monocytes-derived HSCs and rat BM-MSCs has accelerated homing MSCs into injured pancreatic tissue.展开更多
MiRNAs and macrophages play important roles in renal fibrosis.The exosomes secreted by bone marrow mesenchymal stem cells(BM-MSCs)can alleviate renal fibrosis.What is not clear,however,is whether a type of miRNAs in t...MiRNAs and macrophages play important roles in renal fibrosis.The exosomes secreted by bone marrow mesenchymal stem cells(BM-MSCs)can alleviate renal fibrosis.What is not clear,however,is whether a type of miRNAs in the BM-MSCs exosomes can alleviate renal fibrosis by modulating macrophage polarization.First,we take a high-throughput sequencing of miRNAs in exosomes of BM-MSCs from chronic kidney disease(CKD)and normal people.Then we used the UUO mouse model and injected exosomes into the tail vein.The macrophages were stimulated with lipopolysaccharide(LPS).MSC-Exo or exosomes from BM-MSCs transfected with miR-93-5p inhibitor(Inhi-Exo)were added to the culture medium.The macrophages were transfected with miR-93-5p inhibitor or miR-93-5p mimic alone.The expression of miR-93-5p in exosomes of CKD patients was significantly decreased compared with normal people and in the LPS-stimulated macrophages and UUO mice kidneys.After stimulation with LPS,the macrophages polarized toward M1 subtype.MSC-Exo or miR-93-5p mimic promoted macrophages from M1 to M2 subtype.Inhi-Exo or miR-93-5p inhibitor blocked the differentiation from M1 to M2 subtype.Significant fibrotic changes occurred in the kidneys of UUO mice,and M1 macrophages were significantly increased.After injecting exosomes into the tail vein of UUO mice,the degree of renal fibrosis was alleviated,the expression of miR-93-5p in the kidney was significantly increased,and the renal macrophages differentiated from M1 to M2 subtype.These results demonstrated that miR-93-5p in the exosomes derived from BM-MSCs can improve renal fibrosis by inducing macrophage differentiation from M1 to M2 subtype.展开更多
Background:Adult stem cells have a remarkable capacity of differentiating into various cell types necessary for tissue and organ regeneration.Multiple studies have focused on the differentiation potential of mesenchym...Background:Adult stem cells have a remarkable capacity of differentiating into various cell types necessary for tissue and organ regeneration.Multiple studies have focused on the differentiation potential of mesenchymal stem cells(MSCs),however little is known about the molecular characteristics of MSCs and their progenies obtained from donors of different ages.In this study,we analyzed publicly available sequencing data obtained from young(~22-year-old,n=8)and older(~65.5-year-old,n=8)donors of MSCs and their differentiated counterparts:osteocytes,chondrocytes and tenocytes.The raw mRNA and small RNA(non-coding RNA)sequencing data was downloaded from NIH BioProjects and systematically analyzed in order to identify uniquely expressed genes in MSC-derived osteocytes,chondrocytes and tenocytes of younger and older people.Results:We identified many commonly up-and downregulated genes are similar in both groups.However,the young group displayed a greater variety of differentially expressed genes in all analyzed MSC-derived cells.This discrepancy in gene expression profiles between younger and older groups may indicate a greater differentiation potential of MSCs isolated from younger donors.miRNA and mRNA integrated analysis showed key miRNAs that regulate mRNAs in both groups from all differentiated lineages.Conclusions:Our analysis provides additional information to previously reported data for identification of MSC markers of plasticity and engraftment.In addition,our data may shed light upon the molecular mechanisms of age-associated musculoskeletal diseases caused by a decreased capacity of MSCs to regenerate the locomotor system in elderly people.展开更多
文摘Investigating the function of combining induced rat monocytes-derived bone marrow-haemopoietic stem cell (rat BM-HSCs) with LPS and rat bone marrow-mesenchymal stem cell (rat BM-MSCs) was to analyze the acceleration of homing process mechanism in injured pancreas. Mononucleated stem cells were isolated from aspirated whole rat BM using ficoll and cultured in α-MEM complete growth medium in 10 cm petridish. After two days, adherent cells after washing twice in petridish were added α-MEM growth medium and then mesenchymal cells were characterized using CD105 marker in third passage and labeled PKH26. Then haemopoietic stem cells (HSCs) were isolated with magnetic beads CD34+ and differentiated in vitro, and then induced monocytes with LPS. Animal experiment used 28 male Wistar rats, and divided them into 4 groups. After transplantation combined, both cells between monocyte derived HSc (mHSCs) and rat BM-MSC were analyzed expression of pair box gen 4 (Pax4), pancreatic and duodenal homeobox (Pdx1), C-peptide using immunohistochemistry, then secretion of insulin and C-peptide analyzed using indirect ELISA. Results showed that the expressions of Pax4, Pdx1, C-peptide found in the surface membrane cell of pancreatic cell, and secreted C-peptide and insulin were shown significant (P < 0.05) in transplanted group 2, 3 and 4, but in group 3 were transplanted with combined cells more dominant than non-combined cells. Conclusions suggested that combining of induced monocytes-derived HSCs and rat BM-MSCs has accelerated homing MSCs into injured pancreatic tissue.
基金This work was supported by Science and Technology Project of Nantong City(MS 22020009).
文摘MiRNAs and macrophages play important roles in renal fibrosis.The exosomes secreted by bone marrow mesenchymal stem cells(BM-MSCs)can alleviate renal fibrosis.What is not clear,however,is whether a type of miRNAs in the BM-MSCs exosomes can alleviate renal fibrosis by modulating macrophage polarization.First,we take a high-throughput sequencing of miRNAs in exosomes of BM-MSCs from chronic kidney disease(CKD)and normal people.Then we used the UUO mouse model and injected exosomes into the tail vein.The macrophages were stimulated with lipopolysaccharide(LPS).MSC-Exo or exosomes from BM-MSCs transfected with miR-93-5p inhibitor(Inhi-Exo)were added to the culture medium.The macrophages were transfected with miR-93-5p inhibitor or miR-93-5p mimic alone.The expression of miR-93-5p in exosomes of CKD patients was significantly decreased compared with normal people and in the LPS-stimulated macrophages and UUO mice kidneys.After stimulation with LPS,the macrophages polarized toward M1 subtype.MSC-Exo or miR-93-5p mimic promoted macrophages from M1 to M2 subtype.Inhi-Exo or miR-93-5p inhibitor blocked the differentiation from M1 to M2 subtype.Significant fibrotic changes occurred in the kidneys of UUO mice,and M1 macrophages were significantly increased.After injecting exosomes into the tail vein of UUO mice,the degree of renal fibrosis was alleviated,the expression of miR-93-5p in the kidney was significantly increased,and the renal macrophages differentiated from M1 to M2 subtype.These results demonstrated that miR-93-5p in the exosomes derived from BM-MSCs can improve renal fibrosis by inducing macrophage differentiation from M1 to M2 subtype.
文摘Background:Adult stem cells have a remarkable capacity of differentiating into various cell types necessary for tissue and organ regeneration.Multiple studies have focused on the differentiation potential of mesenchymal stem cells(MSCs),however little is known about the molecular characteristics of MSCs and their progenies obtained from donors of different ages.In this study,we analyzed publicly available sequencing data obtained from young(~22-year-old,n=8)and older(~65.5-year-old,n=8)donors of MSCs and their differentiated counterparts:osteocytes,chondrocytes and tenocytes.The raw mRNA and small RNA(non-coding RNA)sequencing data was downloaded from NIH BioProjects and systematically analyzed in order to identify uniquely expressed genes in MSC-derived osteocytes,chondrocytes and tenocytes of younger and older people.Results:We identified many commonly up-and downregulated genes are similar in both groups.However,the young group displayed a greater variety of differentially expressed genes in all analyzed MSC-derived cells.This discrepancy in gene expression profiles between younger and older groups may indicate a greater differentiation potential of MSCs isolated from younger donors.miRNA and mRNA integrated analysis showed key miRNAs that regulate mRNAs in both groups from all differentiated lineages.Conclusions:Our analysis provides additional information to previously reported data for identification of MSC markers of plasticity and engraftment.In addition,our data may shed light upon the molecular mechanisms of age-associated musculoskeletal diseases caused by a decreased capacity of MSCs to regenerate the locomotor system in elderly people.
