Mesenchymal stromal cells (MSCs) can be obtained from several sources and the significant differences in their properties make it crucial to investigate the differentiation potential of MSCs from different sources to ...Mesenchymal stromal cells (MSCs) can be obtained from several sources and the significant differences in their properties make it crucial to investigate the differentiation potential of MSCs from different sources to determine the optimal source of MSCs. We investigated if this biological heterogeneity in MSCs from different sources results in different mechanisms for their differentiation. In this study, we compared the gene expression patterns of phenotypically defined MSCs derived from three ontogenically different sources: Embryonic stem cells (hES-MSCs), Fetal limb (Flb-MSCs) and Bone Marrow (BM-MSCs). Differentially expressed genes between differentiated cells and undifferentiated controls were compared across the three MSC sources. We found minimal overlap (5% - 16%) in differentially expressed gene sets among the three sources. Flb-MSCs were similar to BM-MSCs based on differential gene expression patterns. Pathway analysis of the differentially expressed genes using Ingenuity Pathway Analysis (IPA) revealed a large variation in the canonical pathways leading to MSC differentiation. The similar canonical pathways among the three sources were lineage specific. The Flb-MSCs showed maximum overlap of canonical pathways with the BM-MSCs, indicating that the Flb-MSCs are an intermediate source between the less specialised hES-MSC source and the more specialised BM-MSC source. The source specific pathways prove that MSCs from the three ontogenically different sources use different biological pathways to obtain similar differentiation outcomes. Thus our study advocates the understanding of biological pathways to obtain optimal sources of MSCs for various clinical applications.展开更多
Background:Previous studies have reported that mitochondrial dysfunction participates in the pathological process of osteoarthritis(OA).However,studies that improve mitochondrial function are rare in OA.Mitochondrial ...Background:Previous studies have reported that mitochondrial dysfunction participates in the pathological process of osteoarthritis(OA).However,studies that improve mitochondrial function are rare in OA.Mitochondrial transfer from mesenchymal stem cells(MSCs)to OA chondrocytes might be a cell-based therapy for the improvement of mitochondrial function to prevent cartilage degeneration.This study aimed to determine whether MSCs can donate mitochondria and protect the mitochondrial function and therefore reduce cartilage degeneration.Methods:Bone-marrow-derived mesenchymal stromal cells(BM-MSCs)were harvested from the marrow cavities of femurs and tibia in young rats.OA chondrocytes were gathered from the femoral and tibial plateau in old OA model rats.BM-MSCs and OA chondrocytes were co-cultured and mitochondrial transfer from BM-MSCs to chondrocytes was identified.Chondrocytes with mitochondria transferred from BM-MSCs were selected by fluorescence-activated cell sorting.Mitochondrial function of these cells,including mitochondrial membrane potential(Δψm),the activity of mitochondrial respiratory chain(MRC)enzymes,and adenosine triphosphate(ATP)content were quantified and compared to OA chondrocytes without mitochondrial transfer.Chondrocytes proliferation,apoptosis,and secretion ability were also analyzed between the two groups.Results:Mitochondrial transfer was found from BM-MSCs to OA chondrocytes.Chondrocytes with mitochondrial from MSCs(MSCs+OA group)showed increased mitochondrial membrane potential compared with OA chondrocytes without mitochondria transfer(OA group)(1.79±0.19 vs.0.71±0.12,t=10.42,P<0.0001).The activity of MRC enzymes,including MRC complex I,II,III,and citrate synthase was also improved(P<0.05).The content of ATP in MSCs+OA group was significantly higher than that in OA group(161.90±13.49 vs.87.62±11.07 nmol/mg,t=8.515,P<0.0001).Meanwhile,we observed decreased cell apoptosis(7.09%±0.68%vs.15.89%±1.30%,t=13.39,P<0.0001)and increased relative secretion of type II collagen(2.01±0.14 v展开更多
间充质干细胞具有高度增殖、自我更新和多向分化的潜能。大电导钙离子激活的钾通道M亚族α亚基(potassium large conductance calcium-activated channel,subfamily M,alpha member 1,KCNMA1)介导细胞内K+的外流,使细胞膜超极化,降低细...间充质干细胞具有高度增殖、自我更新和多向分化的潜能。大电导钙离子激活的钾通道M亚族α亚基(potassium large conductance calcium-activated channel,subfamily M,alpha member 1,KCNMA1)介导细胞内K+的外流,使细胞膜超极化,降低细胞的兴奋性。该研究通过制备KCNMA1重组慢病毒载体和空白对照慢病毒载体,将其转染至间充质干细胞内,测定转染复数值(MOI),并通过RT-PCR和Western blot比较转染前后KCNMA1的表达变化情况,检测转染前后细胞微环境中电解质浓度变化。结果成功包装了KCNMA1慢病毒载体和空白病毒载体并转染入干细胞内;含有目的基因的慢病毒转染间充质干细胞后,RT-PCR和Western blot提示KCNMA1过表达,且细胞微环境中K+浓度升高。证实成功地将含KCNMA1的慢病毒载体转染进入大鼠间充质干细胞内,并在细胞内过表达且发挥功能,为体内研究KCNMA1结合干细胞治疗相关疾病奠定了基础。展开更多
This study explored the novel strategy of hypoxic preconditioning of Bone Marrow Mesenchymal Stem Cells (BM-MSCs) before intra vitreal transplantation to improve neuroprotective effects of Retinal Ganglion Cells (RGCs...This study explored the novel strategy of hypoxic preconditioning of Bone Marrow Mesenchymal Stem Cells (BM-MSCs) before intra vitreal transplantation to improve neuroprotective effects of Retinal Ganglion Cells (RGCs) in Acute Glaucoma Models. The methods of this research were isolated mesenchymal stem cells from the bone marrow of adult wild-type Sprague-Dawley (SD) rats. BM-MSCs were cultured under normoxic or hypoxic (1% oxygen for 24 hours) conditions. Normoxic or hypoxic BM-MSCs were transplanted intravitreally 1 week after ocular hypertension induction by acutely increasing IOP to 100 - 120 mmHg for 60 minutes. Rats were killed 4 weeks after transplanted. Apoptosis was examined by tunnel assay and expression Brn3b (Brn3b = RGCs marker) by immunohistochemical analysis of the retina. Results showed that transplantation of hypoxic preconditioning BM-MSCs in acute glaucoma models resulted in a significant apoptosis decreasing (p < 0.05) and an significant increasing in RGCs (p < 0.05), as well as enhanced mor-phologic and functional benefits of stem cell therapy versus normoxic BM-MSCs transplantation. Conclusions: Hypoxic preconditioning enhances the capacity of BM-MSCs transplantation to improve neuroprotective effects of RGCs in Acute Glaucoma Models.展开更多
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
文摘Mesenchymal stromal cells (MSCs) can be obtained from several sources and the significant differences in their properties make it crucial to investigate the differentiation potential of MSCs from different sources to determine the optimal source of MSCs. We investigated if this biological heterogeneity in MSCs from different sources results in different mechanisms for their differentiation. In this study, we compared the gene expression patterns of phenotypically defined MSCs derived from three ontogenically different sources: Embryonic stem cells (hES-MSCs), Fetal limb (Flb-MSCs) and Bone Marrow (BM-MSCs). Differentially expressed genes between differentiated cells and undifferentiated controls were compared across the three MSC sources. We found minimal overlap (5% - 16%) in differentially expressed gene sets among the three sources. Flb-MSCs were similar to BM-MSCs based on differential gene expression patterns. Pathway analysis of the differentially expressed genes using Ingenuity Pathway Analysis (IPA) revealed a large variation in the canonical pathways leading to MSC differentiation. The similar canonical pathways among the three sources were lineage specific. The Flb-MSCs showed maximum overlap of canonical pathways with the BM-MSCs, indicating that the Flb-MSCs are an intermediate source between the less specialised hES-MSC source and the more specialised BM-MSC source. The source specific pathways prove that MSCs from the three ontogenically different sources use different biological pathways to obtain similar differentiation outcomes. Thus our study advocates the understanding of biological pathways to obtain optimal sources of MSCs for various clinical applications.
基金supported by a grant from the Scientific Research Seed Fund of Peking University First Hospital(No.2018SF020)。
文摘Background:Previous studies have reported that mitochondrial dysfunction participates in the pathological process of osteoarthritis(OA).However,studies that improve mitochondrial function are rare in OA.Mitochondrial transfer from mesenchymal stem cells(MSCs)to OA chondrocytes might be a cell-based therapy for the improvement of mitochondrial function to prevent cartilage degeneration.This study aimed to determine whether MSCs can donate mitochondria and protect the mitochondrial function and therefore reduce cartilage degeneration.Methods:Bone-marrow-derived mesenchymal stromal cells(BM-MSCs)were harvested from the marrow cavities of femurs and tibia in young rats.OA chondrocytes were gathered from the femoral and tibial plateau in old OA model rats.BM-MSCs and OA chondrocytes were co-cultured and mitochondrial transfer from BM-MSCs to chondrocytes was identified.Chondrocytes with mitochondria transferred from BM-MSCs were selected by fluorescence-activated cell sorting.Mitochondrial function of these cells,including mitochondrial membrane potential(Δψm),the activity of mitochondrial respiratory chain(MRC)enzymes,and adenosine triphosphate(ATP)content were quantified and compared to OA chondrocytes without mitochondrial transfer.Chondrocytes proliferation,apoptosis,and secretion ability were also analyzed between the two groups.Results:Mitochondrial transfer was found from BM-MSCs to OA chondrocytes.Chondrocytes with mitochondrial from MSCs(MSCs+OA group)showed increased mitochondrial membrane potential compared with OA chondrocytes without mitochondria transfer(OA group)(1.79±0.19 vs.0.71±0.12,t=10.42,P<0.0001).The activity of MRC enzymes,including MRC complex I,II,III,and citrate synthase was also improved(P<0.05).The content of ATP in MSCs+OA group was significantly higher than that in OA group(161.90±13.49 vs.87.62±11.07 nmol/mg,t=8.515,P<0.0001).Meanwhile,we observed decreased cell apoptosis(7.09%±0.68%vs.15.89%±1.30%,t=13.39,P<0.0001)and increased relative secretion of type II collagen(2.01±0.14 v
文摘间充质干细胞具有高度增殖、自我更新和多向分化的潜能。大电导钙离子激活的钾通道M亚族α亚基(potassium large conductance calcium-activated channel,subfamily M,alpha member 1,KCNMA1)介导细胞内K+的外流,使细胞膜超极化,降低细胞的兴奋性。该研究通过制备KCNMA1重组慢病毒载体和空白对照慢病毒载体,将其转染至间充质干细胞内,测定转染复数值(MOI),并通过RT-PCR和Western blot比较转染前后KCNMA1的表达变化情况,检测转染前后细胞微环境中电解质浓度变化。结果成功包装了KCNMA1慢病毒载体和空白病毒载体并转染入干细胞内;含有目的基因的慢病毒转染间充质干细胞后,RT-PCR和Western blot提示KCNMA1过表达,且细胞微环境中K+浓度升高。证实成功地将含KCNMA1的慢病毒载体转染进入大鼠间充质干细胞内,并在细胞内过表达且发挥功能,为体内研究KCNMA1结合干细胞治疗相关疾病奠定了基础。
文摘This study explored the novel strategy of hypoxic preconditioning of Bone Marrow Mesenchymal Stem Cells (BM-MSCs) before intra vitreal transplantation to improve neuroprotective effects of Retinal Ganglion Cells (RGCs) in Acute Glaucoma Models. The methods of this research were isolated mesenchymal stem cells from the bone marrow of adult wild-type Sprague-Dawley (SD) rats. BM-MSCs were cultured under normoxic or hypoxic (1% oxygen for 24 hours) conditions. Normoxic or hypoxic BM-MSCs were transplanted intravitreally 1 week after ocular hypertension induction by acutely increasing IOP to 100 - 120 mmHg for 60 minutes. Rats were killed 4 weeks after transplanted. Apoptosis was examined by tunnel assay and expression Brn3b (Brn3b = RGCs marker) by immunohistochemical analysis of the retina. Results showed that transplantation of hypoxic preconditioning BM-MSCs in acute glaucoma models resulted in a significant apoptosis decreasing (p < 0.05) and an significant increasing in RGCs (p < 0.05), as well as enhanced mor-phologic and functional benefits of stem cell therapy versus normoxic BM-MSCs transplantation. Conclusions: Hypoxic preconditioning enhances the capacity of BM-MSCs transplantation to improve neuroprotective effects of RGCs in Acute Glaucoma Models.
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.
