Critical-sized craniofacial defect repair represents a significant challenge to reconstructive surgeons.Many strategies have been employed in an effort to achieve both a functionally and cosmetically acceptable outcom...Critical-sized craniofacial defect repair represents a significant challenge to reconstructive surgeons.Many strategies have been employed in an effort to achieve both a functionally and cosmetically acceptable outcome.Bone morphogenetic proteins(BMPs)provide a robust osteoinductive cue to stimulate bony growth and remodeling.Previous studies have suggested that the BMP-9 isoform is particularly effective in promoting osteogenic differentiation of mesenchymal progenitor cells.The aim of this study is to characterize the osteogenic capacity of BMP-9 on calvarial mesenchymal progenitor cell differentiation.Reversibly immortalized murine calvarial progenitor cells(iCALs)were infected with adenoviral vectors encoding BMP-9 or GFP and assessed for early and late stages of osteogenic differentiation in vitro and for osteogenic differentiation via in vivo stem cell implantation studies.Significant elevations in alkaline phosphatase(ALP)activity,osteocalcin(OCN)mRNA transcription,osteopontin(OPN)protein expression,and matrix mineralization were detected in BMP-treated cells compared to control.Specifically,ALP activity was elevated on days 3,7,9,11,and 13 post-infection and OCN mRNA expression was elevated on days 8,10,and 14 in treated cells.Additionally,treatment groups demonstrated increased OPN protein expression on day 10 and matrix mineralization on day 14 post-infection relative to control groups.BMP-9 also facilitated the formation of new bone in vivo as detailed by gross,microcomputed tomography,and histological analyses.Therefore,we concluded that BMP-9 significantly stimulates osteogenic differentiation in iCALs,and should be considered an effective agent for calvarial tissue regeneration.展开更多
The feasibility of using cord blood mesenchymal stem/progenitor cells (CB-MSPCs) to regenerate cardiomyocytes and the optimal inducing conditions were investigated. The CB mononuclear cells were cultured in low serum ...The feasibility of using cord blood mesenchymal stem/progenitor cells (CB-MSPCs) to regenerate cardiomyocytes and the optimal inducing conditions were investigated. The CB mononuclear cells were cultured in low serum DMEM medium to produce an adherent layer. After expansion, the adherent cells were added into cardiomyocyte inducing medium supplemented with 5-azacytidine. Cardiogenic specific contractile protein troponin T staining was performed to identify the cardiomy-ocyte-like cells. The results showed that the frequency of CB-MSPCs clones in CB mononuclear cells was 0. 5×10-6 and about 1. 3×107-fold expansion was achieved within 20 sub-cultivation. After car-diogenic induction, 70 % CB-MSPCs was differentiated into cardiomyocyte-like cells. It was indicated that low serum culture could expand CB-MSPCs extensively and the expanded CB-MSPCs could be induced to differentiate into cardiomyocyte-like cells in high efficiency.展开更多
Ovarian mesenchymal cells(oMCs)constitute a distinct microenvironment that supports folliculogenesis under physiological conditions.Supplementation of exogenous non-ovarian mesenchymal-related cells has been reported ...Ovarian mesenchymal cells(oMCs)constitute a distinct microenvironment that supports folliculogenesis under physiological conditions.Supplementation of exogenous non-ovarian mesenchymal-related cells has been reported to be an efficient approach to improve ovarian functions.However,the development and cellular and molecular characteristics of endogenous oMCs remain largely unexplored.In this study,we surveyed the single-cell transcriptomic landscape to dissect the cellular and molecular changes associated with the aging of oMCs in mice.Our results showed that the oMCs were composed of five ovarian differentiatedMC(odMC)populations and one ovarian mesenchymal progenitor(oMP)cell population.These cells could differentiate into various odMCs via an oMP-derived route to construct the ovarian stroma structures.Comparative analysis revealed that ovarian aging was associated with decreased quantity of oMP cells and reduced quality of odMCs.Based on the findings of bioinformatics analysis,we designed different strategies involving supplementation with young oMCs to examine their effects on female fertility and health.Our functional investigations revealed that oMCs supplementation prior to ovarian senescence was the optimal method to improve female fertility and extend the reproductive lifespan of aged females in the longterm.展开更多
基金This work was funded by the Chicago Biomedical Consortium with support from the Searle Funds at The Chicago Community TrustThe reported work was also supported in part by an NIH/NIDCK K08 Career Development Award(#1K08DE020140-01,RRR)This work was also supported in part by The University of Chicago Core Facility Subsidy grant from the National Center for Advancing Translational Sciences(NCATS)of the National Institutes of Health through Grant UL1 TR000430(RRR).
文摘Critical-sized craniofacial defect repair represents a significant challenge to reconstructive surgeons.Many strategies have been employed in an effort to achieve both a functionally and cosmetically acceptable outcome.Bone morphogenetic proteins(BMPs)provide a robust osteoinductive cue to stimulate bony growth and remodeling.Previous studies have suggested that the BMP-9 isoform is particularly effective in promoting osteogenic differentiation of mesenchymal progenitor cells.The aim of this study is to characterize the osteogenic capacity of BMP-9 on calvarial mesenchymal progenitor cell differentiation.Reversibly immortalized murine calvarial progenitor cells(iCALs)were infected with adenoviral vectors encoding BMP-9 or GFP and assessed for early and late stages of osteogenic differentiation in vitro and for osteogenic differentiation via in vivo stem cell implantation studies.Significant elevations in alkaline phosphatase(ALP)activity,osteocalcin(OCN)mRNA transcription,osteopontin(OPN)protein expression,and matrix mineralization were detected in BMP-treated cells compared to control.Specifically,ALP activity was elevated on days 3,7,9,11,and 13 post-infection and OCN mRNA expression was elevated on days 8,10,and 14 in treated cells.Additionally,treatment groups demonstrated increased OPN protein expression on day 10 and matrix mineralization on day 14 post-infection relative to control groups.BMP-9 also facilitated the formation of new bone in vivo as detailed by gross,microcomputed tomography,and histological analyses.Therefore,we concluded that BMP-9 significantly stimulates osteogenic differentiation in iCALs,and should be considered an effective agent for calvarial tissue regeneration.
文摘The feasibility of using cord blood mesenchymal stem/progenitor cells (CB-MSPCs) to regenerate cardiomyocytes and the optimal inducing conditions were investigated. The CB mononuclear cells were cultured in low serum DMEM medium to produce an adherent layer. After expansion, the adherent cells were added into cardiomyocyte inducing medium supplemented with 5-azacytidine. Cardiogenic specific contractile protein troponin T staining was performed to identify the cardiomy-ocyte-like cells. The results showed that the frequency of CB-MSPCs clones in CB mononuclear cells was 0. 5×10-6 and about 1. 3×107-fold expansion was achieved within 20 sub-cultivation. After car-diogenic induction, 70 % CB-MSPCs was differentiated into cardiomyocyte-like cells. It was indicated that low serum culture could expand CB-MSPCs extensively and the expanded CB-MSPCs could be induced to differentiate into cardiomyocyte-like cells in high efficiency.
基金supported by the National Key Research and Development Program of China (2022YFC2703800)the National Natural Science Foundation of China (82230051)the 2115 Talent Development Program of China Agricultural University (1021-00109022)。
文摘Ovarian mesenchymal cells(oMCs)constitute a distinct microenvironment that supports folliculogenesis under physiological conditions.Supplementation of exogenous non-ovarian mesenchymal-related cells has been reported to be an efficient approach to improve ovarian functions.However,the development and cellular and molecular characteristics of endogenous oMCs remain largely unexplored.In this study,we surveyed the single-cell transcriptomic landscape to dissect the cellular and molecular changes associated with the aging of oMCs in mice.Our results showed that the oMCs were composed of five ovarian differentiatedMC(odMC)populations and one ovarian mesenchymal progenitor(oMP)cell population.These cells could differentiate into various odMCs via an oMP-derived route to construct the ovarian stroma structures.Comparative analysis revealed that ovarian aging was associated with decreased quantity of oMP cells and reduced quality of odMCs.Based on the findings of bioinformatics analysis,we designed different strategies involving supplementation with young oMCs to examine their effects on female fertility and health.Our functional investigations revealed that oMCs supplementation prior to ovarian senescence was the optimal method to improve female fertility and extend the reproductive lifespan of aged females in the longterm.
