Osteoarthritis(OA)is a degenerative multifactorial disease with concomitant structural,inflammatory,and metabolic changes that fluctuate in a temporal and patient-specific manner.This complexity has contributed to ref...Osteoarthritis(OA)is a degenerative multifactorial disease with concomitant structural,inflammatory,and metabolic changes that fluctuate in a temporal and patient-specific manner.This complexity has contributed to refractory responses to various treatments.MsCs have shown promise as multimodal therapeutics in mitigating OA symptoms and disease progression.Here,we evaluated 15 randomized controlled clinical trials(RCTs)and 11 nonrandomized RCTs using culture-expanded MSCs in the treatment of knee OA,and we found net positive effects of MSCs on mitigating pain and symptoms(improving function in 12/15 RCTs relative to baseline and in 11/15 RCTs relative to control groups at study endpoints)and on cartilage protection and/or repair(18/21 clinical studies).We examined MsC dose,tissue of origin,and autologous vs.allogeneic origins as well as patient clinical phenotype,endotype,age,sex and level of OA severity as key parameters in parsing MSC clinical effectiveness.The relatively small sample size of 610 patients limited the drawing of definitive conclusions.Nonetheless,we noted trends toward moderate to higher doses of MsCs in select OA patient clinical phenotypes mitigating pain and leading to structural improvements or cartilage preservation.Evidence from preclinical studies is supportive of MsC anti-inflammatory and immunomodulatory effects,but additional investigations on immunomodulatory,chondroprotective and other clinical mechanisms of action are needed.We hypothesize that MsC basal immunomodulatory"fitness"correlates with OA treatment efficacy,but this hypothesis needs to be validated in future studies.We conclude with a roadmap articulating the need to match an OA patient subset defined by molecular endotype and clinical phenotype with basally immunomodulatory"ft"or engineered-to-be-fit-for-OA MsCs in well-designed,data-intensive clinical trials to advance the field.展开更多
Liver fibrosis is a significant health burden,marked by the consistent deposition of collagen.Unfortunately,the currently available treatment approaches for this condition are far from optimal.Lysyl oxidase-like prote...Liver fibrosis is a significant health burden,marked by the consistent deposition of collagen.Unfortunately,the currently available treatment approaches for this condition are far from optimal.Lysyl oxidase-like protein 2(LOXL2)secreted by hepatic stellate cells(HSCs)is a crucial player in the cross-linking of matrix collagen and is a significant target for treating liver fibrosis.Mesenchymal stem cell-derived small extracellular vesicles(MSC-sEVs)have been proposed as a potential treatment option for chronic liver disorders.Previous studies have found that MSC-sEV can be used for microRNA delivery into target cells or tissues.It is currently unclear whether microRNA-4465(miR-4465)can target LOXL2 and inhibit HSC activation.Additionally,it is uncertain whether MSC-sEV can be utilized as a gene therapy vector to carry miR-4465 and effectively inhibit the progression of liver fibrosis.This study explored the effect of miR-4465-modified MSC-sEV(MSC-sEVmiR-4465)on LOXL2 expression and liver fibrosis development.The results showed that miR-4465 can bind specifically to the promoter of the LOXL2 gene in HSC.Moreover,MSC-sEVmiR-4465 inhibited HSC activation and collagen expression by downregulating LOXL2 expression in vitro.MSC-sEVmiR-4465 injection could reduce HSC activation and collagen deposition in the CCl4-induced mouse model.MSC-sEVmiR-4465 mediating via LOXL2 also hindered the migration and invasion of HepG2 cells.In conclusion,we found that MSC-sEV can deliver miR-4465 into HSC to alleviate liver fibrosis via altering LOXL2,which might provide a promising therapeutic strategy for liver diseases.展开更多
Identification of mouse cell lines with properties of primary multipotential mesenchymal stromal cells (MSC) is required to facilitate the use of mouse models for evaluation of mechanisms in bone formation, hematopoie...Identification of mouse cell lines with properties of primary multipotential mesenchymal stromal cells (MSC) is required to facilitate the use of mouse models for evaluation of mechanisms in bone formation, hematopoiesis and cellular therapies for regenerative medicine. Primary murine MSC vary between strains, are difficult to grow in vitro and have inconsistent properties. The main aim of the study was to establish OMA-AD cells as an appropriate model system to conduct studies on MSC, bone formation and hematopoiesis. OMA-AD cells were isolated by differential trypsinization of C57BL/6J mouse bone marrow (BM) cells. The cells were then repassaged, cloned and characterized. OMA-AD cells were immortal and non-tumorigenic, differentiated readily to all mesenchymal cell types including bone, supported mouse and human hematopoiesis and were immunosuppressive. Our results demonstrated that OMA-AD cells possessed the properties of primary MSC. In addition, these cells grew readily and consistently, thereby facilitating future studies of bone formation, hematopoiesis and mesenchymal cells for regenerative medicine.展开更多
The etiology of inflammatory bowel disease(IBD)is multifaceted,involving genetic susceptibility,immune dysregulation,alterations in the gut microbiota,and environmental factors.Both intrinsic and extrinsic factors can...The etiology of inflammatory bowel disease(IBD)is multifaceted,involving genetic susceptibility,immune dysregulation,alterations in the gut microbiota,and environmental factors.Both intrinsic and extrinsic factors can disrupt the intestinal mucosal barrier,leading to chronic nonspecific inflammation,local structural changes,and gastrointestinal dysfunction.Historically,due to a lack of effective treatments,recurrent inflammation and microcirculatory disturbances could result in complications such as intestinal fistulas,strictures,obstructions,perforations,gastrointestinal bleeding,sepsis,etc.,thereby increasing the risk of intestinal cell carcinoma and mortality.While the overall incidence of IBD remains at 0.5%in North America and Europe,its annual incidence is increasing in Asia,Africa,and South America,resulting in a growing number of patients and warning significant attention.Recent research has highlighted mesenchymal stem cell(MSC)therapy as an innovative treatment option for IBD due to its capacity to modulate inflammatory immune responses and promote tissue regeneration.A current preclinical study has shown a promising result,with systemic administration of MSCs in patients with reduced intestinal inflammation and no intestinal inflammation.In addition,in a new study,the use of mesenchymal stem cell-derived exosomes(MSC-EXOs)was successful,a type derived from mesenchymal stem cells,was successful,especially in patients with refractory anal fistula.Consequently,MSC therapy has become a preferred approach in IBD treatment,showcasing the potential application prospects for stem cell-based therapy in IBD.However,clinical research in this field still needs to refine strategies and further explore to lay a solid foundation.展开更多
为了探讨小鼠胚胎主动脉-性腺-中肾(aorta-gonad-mesonephros,AGM)区基质微环境对胚胎造血的作用,分离鉴定小鼠背主动脉(dorsalaorta,DA)来源间充质干细胞样基质细胞(mesenchymal stem cell like stromal cells,MSC like stromal cells...为了探讨小鼠胚胎主动脉-性腺-中肾(aorta-gonad-mesonephros,AGM)区基质微环境对胚胎造血的作用,分离鉴定小鼠背主动脉(dorsalaorta,DA)来源间充质干细胞样基质细胞(mesenchymal stem cell like stromal cells,MSC like stromal cells)并研究其生长特性、表面标志和间质分化能力。取E11.5小鼠胚胎,分离获得DA区细胞,转染质粒pSV3neo-SV40,筛选具有G418抗性的阳性细胞,挑取成纤维样细胞,传代培养并检测其增殖能力,应用流式细胞术检测相关表面标志,诱导其向脂肪、成骨和成软骨细胞分化。结果表明:筛选获得的20个细胞克隆多为成纤维样细胞。mDAF3和mDAF18可在体外传代50代以上,稳定表达G418抗性,细胞倍增时间约为24小时,具有向成骨细胞、脂肪细胞和成软骨细胞分化的能力,流式细胞术检测结果显示其表达CD29、CD44、CD105、Sca-1,弱表达CD34,CD45和CD31。结论:永生化的mDAF3和mDAF18具有MSC的表型特征和分化功能,提示小鼠胚胎DA区可能存在MSC,小鼠胚胎DA区MSC样基质细胞可为研究基质微环境对胚胎造血发育的调控作用提供支持细胞。展开更多
AIM:To explore the secretome efficacy in tumor necrosis factor(TNF)-αstimulated mouse mesenchymal stem cells(MSCs)in a murine model of corneal limbal alkali injury.METHODS:Corneal limbal stem cell deficiency(LSCD)was...AIM:To explore the secretome efficacy in tumor necrosis factor(TNF)-αstimulated mouse mesenchymal stem cells(MSCs)in a murine model of corneal limbal alkali injury.METHODS:Corneal limbal stem cell deficiency(LSCD)was created in the eyes of male C57 mice.Concentrated conditioned medium from TNF-αstimulated MSCs(MSC-CMT)was applied topically for 4 wk,with basal medium and conditioned medium from MSCs as controls.Corneal opacification,corneal inflammatory response,and corneal neovascularization(NV)were evaluated.Corneal epithelial cell apoptosis,corneal conjunctivation,and inflammatory cell infiltration were assessed with TUNEL staining,CK3 and Muc-5 AC immunostaining,and CD11 b immunofluorescence staining,respectively.The effect of TSG-6 was further evaluated by knockdown with short hairpin RNA(sh RNA).RESULTS:Compared to the controls,topical administration of MSC-CMT significantly ameliorated the clinical symptoms of alkali-induced LSCD,with restrained corneal NV,reduced corneal epithelial cell apoptosis,and inhibition of corneal conjunctivation.In addition,MSC-CMT treatment significantly reduced CD11 b+inflammatory cell infiltration,and inhibited the expression of pro-inflammatory cytokines(IL-1β,TNF-αand IL-6).Furthermore,the promotion of corneal epithelial reconstruction by MSC-CMT was largely abolished by TSG-6 knockdown.CONCLUSION:Our study provides evidence that MSCCMT enhances the alleviation of corneal alkali injuries,partially through TSG-6-mediated anti-inflammatory protective mechanisms.MSC-CMT may serve as a potential strategy for treating corneal disorders.展开更多
基金funded by the Canadian Institutes of Health Research(CIHR)(PJT-166089)the Natural Sciences and Engineering Research Council of Canada(NSERC)(RGPIN-2018-05737)supported by the Schroeder Arthritis Institute via the Toronto General and Western Hospital Foundation(University Health Network).
文摘Osteoarthritis(OA)is a degenerative multifactorial disease with concomitant structural,inflammatory,and metabolic changes that fluctuate in a temporal and patient-specific manner.This complexity has contributed to refractory responses to various treatments.MsCs have shown promise as multimodal therapeutics in mitigating OA symptoms and disease progression.Here,we evaluated 15 randomized controlled clinical trials(RCTs)and 11 nonrandomized RCTs using culture-expanded MSCs in the treatment of knee OA,and we found net positive effects of MSCs on mitigating pain and symptoms(improving function in 12/15 RCTs relative to baseline and in 11/15 RCTs relative to control groups at study endpoints)and on cartilage protection and/or repair(18/21 clinical studies).We examined MsC dose,tissue of origin,and autologous vs.allogeneic origins as well as patient clinical phenotype,endotype,age,sex and level of OA severity as key parameters in parsing MSC clinical effectiveness.The relatively small sample size of 610 patients limited the drawing of definitive conclusions.Nonetheless,we noted trends toward moderate to higher doses of MsCs in select OA patient clinical phenotypes mitigating pain and leading to structural improvements or cartilage preservation.Evidence from preclinical studies is supportive of MsC anti-inflammatory and immunomodulatory effects,but additional investigations on immunomodulatory,chondroprotective and other clinical mechanisms of action are needed.We hypothesize that MsC basal immunomodulatory"fitness"correlates with OA treatment efficacy,but this hypothesis needs to be validated in future studies.We conclude with a roadmap articulating the need to match an OA patient subset defined by molecular endotype and clinical phenotype with basally immunomodulatory"ft"or engineered-to-be-fit-for-OA MsCs in well-designed,data-intensive clinical trials to advance the field.
基金supported by the National Natural Science Foundation of China(No.82272421)the Jiangsu Provincial Key Research and Development Program(No.BE2021690)+2 种基金the Changzhou's 14th Five-year Plan Project to Train Highlevel Health Professionals(No.2022CZLJ027)the Scientific Project of Jiangsu Health Commission(No.Z2020038)the Changzhou Sci&Tech Program(No.CJ20220164),China.
文摘Liver fibrosis is a significant health burden,marked by the consistent deposition of collagen.Unfortunately,the currently available treatment approaches for this condition are far from optimal.Lysyl oxidase-like protein 2(LOXL2)secreted by hepatic stellate cells(HSCs)is a crucial player in the cross-linking of matrix collagen and is a significant target for treating liver fibrosis.Mesenchymal stem cell-derived small extracellular vesicles(MSC-sEVs)have been proposed as a potential treatment option for chronic liver disorders.Previous studies have found that MSC-sEV can be used for microRNA delivery into target cells or tissues.It is currently unclear whether microRNA-4465(miR-4465)can target LOXL2 and inhibit HSC activation.Additionally,it is uncertain whether MSC-sEV can be utilized as a gene therapy vector to carry miR-4465 and effectively inhibit the progression of liver fibrosis.This study explored the effect of miR-4465-modified MSC-sEV(MSC-sEVmiR-4465)on LOXL2 expression and liver fibrosis development.The results showed that miR-4465 can bind specifically to the promoter of the LOXL2 gene in HSC.Moreover,MSC-sEVmiR-4465 inhibited HSC activation and collagen expression by downregulating LOXL2 expression in vitro.MSC-sEVmiR-4465 injection could reduce HSC activation and collagen deposition in the CCl4-induced mouse model.MSC-sEVmiR-4465 mediating via LOXL2 also hindered the migration and invasion of HepG2 cells.In conclusion,we found that MSC-sEV can deliver miR-4465 into HSC to alleviate liver fibrosis via altering LOXL2,which might provide a promising therapeutic strategy for liver diseases.
文摘Identification of mouse cell lines with properties of primary multipotential mesenchymal stromal cells (MSC) is required to facilitate the use of mouse models for evaluation of mechanisms in bone formation, hematopoiesis and cellular therapies for regenerative medicine. Primary murine MSC vary between strains, are difficult to grow in vitro and have inconsistent properties. The main aim of the study was to establish OMA-AD cells as an appropriate model system to conduct studies on MSC, bone formation and hematopoiesis. OMA-AD cells were isolated by differential trypsinization of C57BL/6J mouse bone marrow (BM) cells. The cells were then repassaged, cloned and characterized. OMA-AD cells were immortal and non-tumorigenic, differentiated readily to all mesenchymal cell types including bone, supported mouse and human hematopoiesis and were immunosuppressive. Our results demonstrated that OMA-AD cells possessed the properties of primary MSC. In addition, these cells grew readily and consistently, thereby facilitating future studies of bone formation, hematopoiesis and mesenchymal cells for regenerative medicine.
文摘The etiology of inflammatory bowel disease(IBD)is multifaceted,involving genetic susceptibility,immune dysregulation,alterations in the gut microbiota,and environmental factors.Both intrinsic and extrinsic factors can disrupt the intestinal mucosal barrier,leading to chronic nonspecific inflammation,local structural changes,and gastrointestinal dysfunction.Historically,due to a lack of effective treatments,recurrent inflammation and microcirculatory disturbances could result in complications such as intestinal fistulas,strictures,obstructions,perforations,gastrointestinal bleeding,sepsis,etc.,thereby increasing the risk of intestinal cell carcinoma and mortality.While the overall incidence of IBD remains at 0.5%in North America and Europe,its annual incidence is increasing in Asia,Africa,and South America,resulting in a growing number of patients and warning significant attention.Recent research has highlighted mesenchymal stem cell(MSC)therapy as an innovative treatment option for IBD due to its capacity to modulate inflammatory immune responses and promote tissue regeneration.A current preclinical study has shown a promising result,with systemic administration of MSCs in patients with reduced intestinal inflammation and no intestinal inflammation.In addition,in a new study,the use of mesenchymal stem cell-derived exosomes(MSC-EXOs)was successful,a type derived from mesenchymal stem cells,was successful,especially in patients with refractory anal fistula.Consequently,MSC therapy has become a preferred approach in IBD treatment,showcasing the potential application prospects for stem cell-based therapy in IBD.However,clinical research in this field still needs to refine strategies and further explore to lay a solid foundation.
文摘为了探讨小鼠胚胎主动脉-性腺-中肾(aorta-gonad-mesonephros,AGM)区基质微环境对胚胎造血的作用,分离鉴定小鼠背主动脉(dorsalaorta,DA)来源间充质干细胞样基质细胞(mesenchymal stem cell like stromal cells,MSC like stromal cells)并研究其生长特性、表面标志和间质分化能力。取E11.5小鼠胚胎,分离获得DA区细胞,转染质粒pSV3neo-SV40,筛选具有G418抗性的阳性细胞,挑取成纤维样细胞,传代培养并检测其增殖能力,应用流式细胞术检测相关表面标志,诱导其向脂肪、成骨和成软骨细胞分化。结果表明:筛选获得的20个细胞克隆多为成纤维样细胞。mDAF3和mDAF18可在体外传代50代以上,稳定表达G418抗性,细胞倍增时间约为24小时,具有向成骨细胞、脂肪细胞和成软骨细胞分化的能力,流式细胞术检测结果显示其表达CD29、CD44、CD105、Sca-1,弱表达CD34,CD45和CD31。结论:永生化的mDAF3和mDAF18具有MSC的表型特征和分化功能,提示小鼠胚胎DA区可能存在MSC,小鼠胚胎DA区MSC样基质细胞可为研究基质微环境对胚胎造血发育的调控作用提供支持细胞。
基金Supported by the National Natural Science Foundation of China(No.81670829)Shandong Provincial Natural Science Fund(No.ZR2018PH020)。
文摘AIM:To explore the secretome efficacy in tumor necrosis factor(TNF)-αstimulated mouse mesenchymal stem cells(MSCs)in a murine model of corneal limbal alkali injury.METHODS:Corneal limbal stem cell deficiency(LSCD)was created in the eyes of male C57 mice.Concentrated conditioned medium from TNF-αstimulated MSCs(MSC-CMT)was applied topically for 4 wk,with basal medium and conditioned medium from MSCs as controls.Corneal opacification,corneal inflammatory response,and corneal neovascularization(NV)were evaluated.Corneal epithelial cell apoptosis,corneal conjunctivation,and inflammatory cell infiltration were assessed with TUNEL staining,CK3 and Muc-5 AC immunostaining,and CD11 b immunofluorescence staining,respectively.The effect of TSG-6 was further evaluated by knockdown with short hairpin RNA(sh RNA).RESULTS:Compared to the controls,topical administration of MSC-CMT significantly ameliorated the clinical symptoms of alkali-induced LSCD,with restrained corneal NV,reduced corneal epithelial cell apoptosis,and inhibition of corneal conjunctivation.In addition,MSC-CMT treatment significantly reduced CD11 b+inflammatory cell infiltration,and inhibited the expression of pro-inflammatory cytokines(IL-1β,TNF-αand IL-6).Furthermore,the promotion of corneal epithelial reconstruction by MSC-CMT was largely abolished by TSG-6 knockdown.CONCLUSION:Our study provides evidence that MSCCMT enhances the alleviation of corneal alkali injuries,partially through TSG-6-mediated anti-inflammatory protective mechanisms.MSC-CMT may serve as a potential strategy for treating corneal disorders.
