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How mesenchymal stem cell cotransplantation with hematopoietic stem cells can improve engraftment in animal models

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World Journal of stem cells 2022年第8期14卷 658-679页

作者： Murilo Montenegro Garrigós Fernando Anselmo de Oliveira Mariana Penteado Nucci Leopoldo Penteado Nucci Arielly da Hora Alves Olivia Furiama Metropolo Dias Lionel Fernel Gamarra Hospital Israelita Albert Einstein São Paulo 05652-900São PauloBrazil Instituto de Química Universidade de São PauloSão Paulo 05508-900São PauloBrazil LIM44-Hospital das Clínicas Faculdade Medicina da Universidade de São PauloSão Paulo 05403-000Brazil Centro Universitário do Planalto Central Área Especial para Industria nº02 Setor Leste-Gama-DFBrasília 72445-020Distrito FederalBrazil

BACKGROUND Bone marrow transplantation(BMT)can be applied to both hematopoietic and nonhematopoietic diseases;nonetheless,it still comes with a number of challenges and limitations that contribute to treatment failure.Bearing this in mind,a possible way to increase the success rate of BMT would be cotransplantation of mesenchymal stem cells(MSCs)and hematopoietic stem cells(HSCs)to improve the bone marrow niche and secrete molecules that enhance the hematopoietic engraftment.AIM To analyze HSC and MSC characteristics and their interactions through cotransplantation in murine models.METHODS We searched for original articles indexed in PubMed and Scopus during the last decade that used HSC and MSC cotransplantation and in vivo BMT in animal models while evaluating cell engraftment.We excluded in vitro studies or studies that involved graft versus host disease or other hematological diseases and publications in languages other than English.In PubMed,we initially identified 555 articles and after selection,only 12 were chosen.In Scopus,2010 were identified,and six were left after the screening and eligibility process.RESULTS Of the 2565 articles found in the databases,only 18 original studies met the eligibility criteria.HSC distribution by source showed similar ratios,with human umbilical cord blood or animal bone marrow being administered mainly with a dose of 1×10^(7) cells by intravenous or intrabone routes.However,MSCs had a high prevalence of human donors with a variety of sources(umbilical cord blood,bone marrow,tonsil,adipose tissue or fetal lung),using a lower dose,mainly 106 cells and ranging 104 to 1.5×107 cells,utilizing the same routes.MSCs were characterized prior to administration in almost every experiment.The recipient used was mostly immunodeficient mice submitted to low-dose irradiation or chemotherapy.The main technique of engraftment for HSC and MSC cotransplantation evaluation was chimerism,followed by hematopoietic reconstitution and survival analysis.Besides the engraftment,homing and cellular

关键词： mesenchymal stem cells Hematopoietic stem cells Bone marrow transplantation Cotransplantation Hematopoietic reconstitution Engraftment

Application of Cell Penetrating Peptide in Magnetic Resonance Imaging of Bone Marrow mesenchymal stem cells

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Acta Biochimica et Biophysica Sinica 2006年第12期38卷 865-873页

作者： Min LIU~1 You-Min GUO~1 Jun-Le YANG~1 Peng WANG~1 Lin-Yu ZHAO~2 Nian SHEN~2 Si-Cen WANG~3 Xiao-Juan GUO~1 Qi-Fei WU~21 Imaging Center,the 2nd Affiliated Hospital of Medical School,Xi’an Jiaotong University,Xi’an 710004,China 2 Key Laboratory of Environment and Genes Related to Diseases of Ministry of Education,Xi’an Jiaotong University,Xi’an 710061,China 3 Department of Pharmacy,Medical School,Xi’an Jiaotong University,Xi’an 710061,China Imaging Center the 2nd Affiliated Hospital of Medical School Xi'an Jiaotong UniversityXi'an 710004 China Key Laboratory of Environment and Genes Related to Diseases of Ministry of Education Xi'an Jiaotong UniversityXi'an 710061 China Department of Pharmacy Medical School Xi'an Jiaotong UniversityXi'an 710061 China

Tracking the distribution and differentiation of stem cells by high-resolution imaging techniqueswould have significant clinical and research implications.In this study,a model cell-penetrating peptide wasused to carry gadolinium particles for magnetic resonance imaging (MRI) of mesenchymal stem cells (MSCs).MSCs were isolated from rat bone marrow and identified by osteogenic differentiation in vitro.The cell-penetrating peptide labeled with fluorescein-5-isothiocyanate (FITC) and gadolinium was synthesized by asolid-phase peptide synthesis method.Fluorescein imaging analysis confirmed that this new peptide couldinternalize into the cytoplasm and nucleus at room temperature,4℃ and 37℃.Gadolinium were efficientlyinternalized into mesenchymal stem cells by the peptide in a time or concentration-dependent manner,resulting in intercellular shortening of longitudinal relaxation enhancements,which were obviously detectedby 1.5 Tesla Magnetic Resonance Imaging.Cytotoxicity assay and flow cytometric analysis showed thatthe intercellular contrast medium incorporation did not affect cell viability at the tested concentrations.Thein vitro experiment results suggested that the new constructed peptides could be a vector for trackingMSCs.

关键词： mesenchymal stem cells cell-penetrating peptide gadolinium fluorescein magnetic resonance imaging

Human umbilical cord-derived mesenchymal stem cells alleviate oxidative stress-induced islet impairment via the Nrf2/HO-1 axis

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Journal of Molecular Cell Biology 2023年第5期15卷 38-51页

作者： Peng Liu Baige Cao Yang Zhou Huina Zhang Congrong Wang Shanghai Diabetes Institute Department of Endocrinology and MetabolismShanghai Key Laboratory of Diabetes MellitusShanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of MedicineShanghai 200233China Department of Endocrinology&Metabolism Shanghai Fourth People’s HospitalSchool of MedicineTongji UniversityShanghai 200434China Translational Medical Center for stem Cell Therapy Shanghai East HospitalSchool of MedicineTongji UniversityShanghai 200092China stem Cell Translational Research Center Tongji HospitalSchool of MedicineTongji UniversityShanghai 200065China

Hyperglycaemia-induced oxidative stress may disrupt insulin secretion andβ-cell survival in diabetes mellitus by overproducing reactive oxygen species.Human umbilical cord-derived mesenchymal stem cells(hUC-MSCs)exhibit antioxidant properties.However,the mechanisms by which hUC-MSCs protectβ-cells from high glucose-induced oxidative stress remain underexplored.In this study,we showed that intravenously injected hUC-MSCs engrafted into the injured pancreas and promoted pancreaticβ-cell function in a mouse model of type 1 diabetes mellitus.The in vitro study revealed that hUC-MSCs attenuated high glucoseinduced oxidative stress and preventedβ-cell impairment via the Nrf2/HO-1 signalling pathway.Nrf2 knockdown partially blocked the anti-oxidative effect of hUC-MSCs,resulting inβ-cell decompensation in a high-glucose environment.Overall,these findings provide novel insights into how hUC-MSCs protectβ-cells from high glucose-induced oxidative stress.

关键词： type 1 diabetes β-cell protection mesenchymal stem cells oxidative stress Nrf2

Effects of cytokines and chemokines on migration of mesenchymal stem cells following spinal cord injury

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Neural Regeneration Research 2012年第14期7卷 1106-1112页

作者： Longyun Li Maoguang Yang Chunxin Wang Qiheng Zhao Jian Liu Chuanguo Zhan Zhi Liu Xuepeng Li Weihua Wang Xiaoyu Yang Department of Orthopedics China-Japan Friendship HospitalJilin UniversityChangchun 130033Jilin ProvinceChina

We investigated the effects of cytokines and chemokines and their associated signaling pathways on mesenchymal stem cell migration after spinal cord injury, to determine their roles in the curative effects of mesenchymal stem cells. This study reviewed the effects of tumor necrosis factor-α, vascular endothelial growth factor, hepatocyte growth factor, platelet-derived growth factor, basic fibroblast growth factor, insulin like growth factor-I, stromal cell-derived factor and monocyte chemoattractant protein-1, 3 during mesenchymal stem cell migration to damaged sites, and analyzed the signal transduction pathways involved in their effects on mesenchymal stem cell migration. The results confirmed that phosphatidylinositol 3-kinase/serine/threonine protein kinases and nuclear factor-KB play crucial roles in the migration of mesenchymal stem cells induced by cytokines and chemokines.

关键词： spinal cord injury mesenchymal stem cells migration cytokine chemokine signaling pathway neural regeneration

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Impact of source tissue and ex vivo expansion on the characterization of goat mesenchymal stem cells

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Journal of Animal Science and Biotechnology 2015年第2期6卷 230-251页

作者： Nuradilla Mohamad-Fauzi Pablo J Ross Elizabeth A Maga James D Murray Department of Animal Science University of California Institute of Ocean and Earth Sciences University of Malaya Department of Population Health and Reproduction University of California

Background： There is considerable interest in using goats as models for genetically engineering dairy animals and also for using stem cells as therapeutics for bone and cartilage repair. mesenchymal stem cells （MSCs） have been isolated and characterized from various species, but are poorly characterized in goats. Results： Goat MSCs isolated from bone marrow （BM-MSCs） and adipose tissue （ASCs） have the ability to undergo osteogenic, adipogenic and chondrogenic differentiation. Cytochemical staining and gene expression analysis show that ASCs have a greater capacity for adipogenic differentiation compared to BM-MSCs and fibroblasts. Different methods of inducing adipogenesis also affect the extent and profile of adipogenic differentiation in MSCs. Goat fibroblasts were not capable of osteogenesis, hence distinguishing them from the MSCs. Goat MSCs and fibroblasts express CD90, CD105, CD73 but not CD45, and exhibit cytoplasmic localization of OCT4 protein. Goat MSCs can be stably transfected by Nucleofection, but, as evidenced by colony-forming efficiency （CFE）, yield significantly different levels of progenitor cells that are robust enough to proliferate into colonies of integrants following G418 selection. BM-MSCs expanded over increasing passages in vitro maintained karyotypic stability up to 20 passages in culture, exhibited an increase in adipogenic differentiation and CFE, but showed altered morphology and amenability to genetic modification by selection. Conclusions： Our findings provide characterization information on goat MSCs, and show that there can be significant differences between MSCs isolated from different tissues and from within the same tissue. Fibroblasts do not exhibit trilineage differentiation potential at the same capacity as MSCs, making it a more reliable method for distinguishing MSCs from fibroblasts, compared to cell surface marker expression.

关键词： Adipose Bone marrow Characterization Differentiation Goat mesenchymal stem cells

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Differentiation potential of bone marrow mesenchymal stem cells in duck

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Journal of Genetics and Genomics 2009年第3期36卷 133-140页

作者： Linfeng Li Xiujuan Bai Xuelian Gong Hongkun Liu Lina Chen Weijun Guan Yuehui Ma Institute of Animal Science Chinese Academy of Agricultural Science Beijing 100193 China College of Animal Science and Technology Northeast Agricultural University Harbin 150000 China

The bone marrow mesenchymal stem cells （MSCs） are multipotent stem cells which can differentiate into mesenchymal cells in vitro. In this study, MSCs in duck were isolated from bone marrow by density gradient centrifuge separation, purified and expanded in the me- dium. The primary MSCs were expanded for 11 passages. The different-passage MSCs were induced to differentiate into osteoblasts and neuron-like cells. Karyotype analysis indicated that MSCs kept diploid condition and the hereditary feature was stable. The different- passage MSCs expressed CD44, ICAM-1 and SSEA-4, but not CD34, CD45 and SSEA-1 when detected by immunofluorescence staining There was no significant difference among the positive rates of passages 2, 6 and 8 （P 〉 0.05）, but a significant difference existed among those of passages 2, 6, 8 and 11 （P 〈 0.05）. After the osteogenic inducement was added, the induced different-passage MSCs expressed high-level alkaline phosphatase （ALP）, and are positive for tetracycline staining, Alizarin Red staining and Von Kossa staining. After the neural inducement was added, about 70% cells exhibited typical neuron-like phenotype, the induced different-passage MSCs expressed Nestin, neuron-specific enolase （NSE） and glial fibrillary acidic protein （GFAP） when detected by immunofluorescence staining. There was no significant difference among the positive rates of passages 3, 4 and 6 （P〉0.05）, but a significant difference existed among those of passages 3, 4, 6 and 8 （P〈0.05）. These results suggest that MSCs in duck were capable of differentiating into osteoblasts and neuron-like cells in vitro.

关键词： duck mesenchymal stem cells induction differentiation

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MicroRNA-663 induces immune dysregulation by inhibiting TGF-β1 production in bone marrow-derived mesenchymal stem cells in patients with systemic lupus erythematosus

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Cellular & Molecular Immunology 2019年第3期16卷 260-274页

作者： Linyu Geng Xiaojun Tang Kangxing Zhou Dandan Wang Shiying Wang Genhong Yao Weiwei Chen Xiang Gao Wanjun Chen Songtao Shi Nan Shen Xuebing Feng Lingyun Sun Department of Rheumatology and Immunology The Affiliated Drum Tower Hospital of Nanjing University Medical SchoolNanjing 210008China Key Laboratory of Model Animal for Disease Study Model Animal Research CenterNanjing UniversityNanjing 210000China Mucosal Immunology Section OPCBNational Institute of Dental and Craniofacial ResearchNational Institutes of HealthBethesda 20892-2190MDUSA Department of Anatomy and Cell Biology School of Dental MedicineUniversity of PennsylvaniaPhiladelphia 19104-6004PAUSA Joint Molecular Rheumatology Laboratory of the Institute of Health Sciences and Shanghai Renji Hospital Shanghai Institutes for Biological SciencesChinese Academy of Sciencesand Shanghai Jiaotong University School of MedicineShanghaiChina

mesenchymal stem cells(MSCs)are critical for immune regulation.Although several microRNAs(miRNAs)have been shown to participate in autoimmune pathogenesis by affecting lymphocyte development and function,the roles of miRNAs in MSC dysfunction in autoimmune diseases remain unclear.Here,we show that patients with systemic lupus erythematosus(SLE)display a unique miRNA signature in bone marrow-derived MSCs(BMSCs)compared with normal controls,among which miR-663 is closely associated with SLE disease activity.MiR-663 inhibits the proliferation and migration of BMSCs and impairs BMSC-mediated downregulation of follicular T helper(Tfh)cells and upregulation of regulatory T(Treg)cells by targeting transforming growth factorβ1(TGF-β1).MiR-663 overexpression weakens the therapeutic effect of BMSCs,while miR-663 inhibition improves the remission of lupus disease in MRL/lpr mice.Thus,miR-663 is a key mediator of SLE BMSC regulation and may serve as a new therapeutic target for the treatment of lupus.

关键词： immune dysregulation mesenchymal stem cells miR-663 systemic lupus erythematosus transforming growth factorβ1

Neural EGF-like protein 1(NELL-1):Signaling crosstalk in mesenchymal stem cells and applications in regenerative medicine

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Genes & Diseases 2017年第3期4卷 127-137页

作者： Mikhail Pakvasa Alex Alverdy Sami Mostafa Eric Wang Lucy Fu Alexander Li Leonardo Oliveira Aravind Athiviraham Michael J.Lee Jennifer Moriatis Wolf Tong-Chuan He Guillermo A.Ameer Russell R.Reid The University of Chicago Pritzker School of MedicineChicagoIL 60637USA Laboratory of Craniofacial Biology and Development Section of Plastic and Reconstructive SurgeryDepartment of SurgeryThe University of Chicago Medical CenterChicagoIL 60637USA Rosalind Franklin University Chicago Medical SchoolNorth ChicagoIL 60064USA Molecular Oncology Laboratory Department of Orthopaedic Surgery and Rehabilitation MedicineThe University of Chicago Medical CenterChicagoIL 60637USA Department of Biomedical Engineering Northwestern UniversityEvanstonIL 60208USA Center for Advanced Regenerative Engineering Northwestern UniversityEvanstonIL 60208USA Department of Surgery Feinberg School of MedicineNorthwestern UniversityChicagoIL 60611USA

Bone tissue regeneration holds the potential to solve both osteoporosis and large skeletal defects,two problems associated with significant morbidity.The differentiation of mesenchymal stem cells into the osteogenic lineage requires a specific microenvironment and certain osteogenic growth factors.Neural EGF Like-Like molecule 1(NELL-1)is a secreted glycoprotein that has proven,both in vitro and in vivo,to be a potent osteo-inductive factor.Furthermore,it has been shown to repress adipogenic differentiation and inflammation.NELL-1 can work synergistically with other osteogenic factors such as Bone Morphogenic Protein(BMP)2 and9,and has shown promise for use in tissue engineering and as a systemically administered drug for the treatment of osteoporosis.Here we provide a comprehensive up-to-date review on the molecular signaling cascade of NELL-1 in mesenchymal stem cells and potential applications in bone regenerative engineering.

关键词： Bone tissue engineering mesenchymal stem cells NELL-1 NEL-like protein 1 Osteogenic differentiation

Monitoring the source of mesenchymal stem cells in patients after transplantation of mismatched-sex hematopoietic stem cells plus thirdparty cells

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Chinese Medical Journal 2013年第22期126卷 4254-4259页

作者： WANG Jing HUANG Xiao-jun XU Lan-ping LIU Dai-hong CHEN Huan CHEN Yu-hong LAI Yue-yun HOU Rui-qin QIN Xiao-ying LIU Kai-yan Peking University Institute of Hematology Peking University People's Hospital Beijing 100044 China

Background In bone marrow transplant patients, the microenvironment in bone marrow is damaged after chemotherapy or radiotherapy. Subsequent to allogenic hematopoietic stem cell transplantation in patients with clinically successful engraftments, the source of mesenchymal stem cells （MSCs） remains controversial. To further verify the stimulatory effect of the simultaneous transplantation of cells from second donors on engraftment success for hematopoietic stem cell transplantation in support of donor MSCs engraftments, the aim of this study is to monitor the dynamics of the engraftment of bone marrow-derived MSCs in patients after transplantation with mismatched-sex hematopoietic stem and third-party cells. Methods In this study, the hematopoietic stem cells from 32 clinical donors of different sexes that resulted in successful engraftments were selected for transplantation and were classified into three groups for research purposes： group A consisted of 14 cases of transplantation with bone marrow and recruited peripheral hematopoietic stem cell transplantation, group B contained 8 cases of simultaneous re-transfusion of MSCs from the second donor, and group C contained 10 cases of simultaneous re-transfusion of umbilical blood from the second donor. The bone marrow from 32 patients with successful engraftments of hematopoietic transplantation were selected and sub-cultured with MSCs. Flow cytometry （FCM） was used to measure the expression of surface antigens on MSCs. Denaturing high-performance liquid chromatography （DHPLC） in combination with polymerase chain reaction amplification of short tandem repeats （STR- PCR） was used to measure the engraftment status of fifth-generation MSCs in patients. Fluorescence in situ hybridization （FISH） revealed the sex origin of the fifth-generation MSCs in 32 patients. Dynamic examinations were performed on patients receiving donor transplantations. Results The progenies of fifth-generation MSCs were successfully cultured in 32 cases. The results of FCM demonstrated that the

关键词： hematopoietic stem cell transplantation mesenchymal stem cells source of engraftment