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Recent updates on the biological basis of heterogeneity in bone marrow stromal cells/ skeletal stem cells

Biomaterials Translational 2022年第1期3卷 3-16页

作者： Deepika Arora Pamela Gehron Robey National Institute of Dental and Craniofacial Research National Institutes of HealthDepartment of Health and Human ServicesBethesdaMDUSA Biosystems and Biomaterials Division National Institute of Standards and TechnologyDepartment of CommerceGaithersburgMDUSA Department of Biotechnology School of Biological Engineering&Life SciencesShobhit Institute of Engineering&Technology(Deemed-to-be-University)MeerutIndia

Based on studies over the last several decades,the self-renewing skeletal lineages derived from bone marrow stroma could be an ideal source for skeletal tissue engineering.However,the markers for osteogenic precursors;i.e.,bone marrow-derived skeletal stem cells(SSCs),in association with other cells of the marrow stroma(bone marrow stromal cells,BMSCs)and their heterogeneous nature both in vivo and in vitro remain to be clarified.This review aims to highlight:i)the importance of distinguishing BMSCs/SSCs from other“mesenchymal stem/stromal cells”,and ii)factors that are responsible for their heterogeneity,and how these factors impact on the differentiation potential of SSCs towards bone.The prospective role of SSC enrichment,their expansion and its impact on SSC phenotype is explored.Emphasis has also been given to emerging single cell RNA sequencing approaches in scrutinizing the unique population of SSCs within the BMSC population,along with their committed progeny.Understanding the factors involved in heterogeneity may help researchers to improvise their strategies to isolate,characterize and adopt best culture practices and source identification to develop standard operating protocols for developing reproducible stem cells grafts.However,more scientific understanding of the molecular basis of heterogeneity is warranted that may be obtained from the robust high-throughput functional transcriptomics of single cells or clonal populations.

关键词： bone marrow stromal cells clonal analysis heterogeneity single cell analysis skeletal stem cells

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An ectopic study of tissue-engineered bone with Nell-1 gene modified rat bone marrow stromal cells in nude mice

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Chinese Medical Journal 2009年第8期122卷 972-979页

作者： HU Jing-zhou ZHANG Zhi-yuan ZHAO Jun ZHANG Xiu-li LIU Gen-tao JIANG Xin-quan Department of Oral and Maxillofacial Surgery Ninth People's Hospital School of Medicine Shanghai Jiao Tong University Shanghai 200011 China Shanghai Research Institute of Stomatology Shanghai Key Lab of Stomatology Ninth People's Hospital School of Medicine Shanghai Jiao Tong University Shanghai 200011 China Department of Neurosurgery Cedars-Sinai Medical Center Los Angeles CA 90048 USA

Background Tissue engineering techniques combined with gene therapy have been recently used to improve osteogenesis. NEL-like molecule-1 （Nell-1）, a novel growth factor, has been reported to have specificity for osteochondral lineage. The study assessed the osteogenic differentiation of rat bone marrow stromal cells （bMSCs） after Nell-1 gene modification and examined its ectopic bone formation ability in a nude mice model with tissue engineering technique. Methods bMSCs obtained from Fischer 344 rats were transduced with either AdNell-1 （Nell-1 group） or Ad-β-galactosidase （AdLacZ, LacZ group） or left untransduced （untransduced group）. The expression of Nell-1 protein was determined by Western blotting and transfer efficiency was assessed, mRNA expressions of osteopontin （OP）, bone sialoprotein （BSP） and osteocalcin （OC） were assessed by real-time PCR 0, 3, 7, 14, and 21 days after gene transfer. Alkaline phosphatase （ALP） activity was measured and von Kossa test was also conducted. Finally, with a tissue engineering technique, gene transduced bMSCs, combining with β-tricalcium phosphate （β-TCP） at a concentration of 2×10^7 cells/ml, were implanted at subcutaneous sites on the back of nude mice. Four weeks after surgery, the implants were evaluated with histological staining and computerized analysis of new bone formation. Results Under current transduction conditions, gene transfer efficiency reached （57.9±6.8）%. Nell-1 protein was detected in Nell-1 group but not in untransduced group and LacZ group. Induced by Nell-1, BSP and OP expression were increased at intermediate stage and OC expression was increased at later stage. ALP activity and the number of calcium nodules were highest in Nell-1 group. Four weeks after implanted into nude mice subcutaneously, the percentage of new bone area in Nell-1 group was （18.1±5.0）%, significantly higher than those of untransduced group （11.3±3.2）% and LacZ group （12.3±3.1）% （P〈0.05）. Conclusions This study has demonstrated the ability of Nell-1 to induce osteogenic differen

关键词： bone marrow stromal cells Nel-like protein type 1 gene tissue engineering

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Glucocorticoids affect the metabolism of bone marrow stromal cells and lead to osteonecrosis of the femoral head： a review

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Chinese Medical Journal 2012年第1期125卷 134-139页

作者： TAN Gang KANG Peng-de PEI Fu-xing Department of Orthopedic Surgery West China HospitalSichuan UniversityChengduSichuan 610041China

Objective To review the recent developments in the mechanisms of glucocorticoids induced osteonecrosis of femoral head （ONFH） and introduce a new theory of ONFH.Data sources Both Chinese- and English-language literatures were searched using MEDLINE （1997-2011）,Pubmed （1997-2011 ） and the Index of Chinese-language Literature （1997-2011 ）.Study selection Data from published articles about mechanisms of glucocorticoids induced ONFH in recent domestic and foreign literature were selected.Data extraction Data were mainly extracted from 61 articles which are listed in the reference section of this review.Results Glucocorticoids are steroid hormones secreted by the adrenal cortex that play a pivotal role in the regulation of a variety of developmental,metabolic and immune functions.However,high dose of exogenous glucocorticoids usage is the most common non-traumatic cause of ON FH.Glucocorticoids can affect the metabolisms of osteoblasts,osteoclasts,bone marrow stromal cells and adipocytes which decrease osteoblasts formation but increase adipocytes formation and cause ONFH finally.Conclusions Glucocorticoids affect the differentiation of mesenchymal stem cells,through activating or inhibiting the related transcript regulators of osteogenesis and adipogenesis.At last,the size and volume of mesenchymal stem cells derived adipocytes will increase amazingly,but the osteoblasts will be decreased obviously.In the meantime,the activity of the osteoclasts will be activated.So,these mechanisms work together and lead to ONFH.

关键词： glucocorticoids osteonecrosis of femoral head peroxisome proliferator-activated receptor-y Runx2 Dickkopf-1 bone marrow stromal cells

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Possible Mechanism of Therapeutic Effect of 3-Methyl-l-phenyl-2-pyrazolin-5-one and bone marrow stromal cells Combination Treatment in Rat Ischemic Stroke Model

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Chinese Medical Journal 2016年第12期129卷 1471-1476页

作者： Li-Hua Shen Jin Chen Hua-Chao Shen Min Ye Xiao-Fei Liu Wen-Sen Ding Ya-Feng Sheng Xin-Sheng Ding Department of Neurology The Affiliated Hospital of Nantong University Nantong Jiangsu 226001 China The BenQ Neurological Institute of Nanjing Medical University Nanjing Jiangsu 210029 China Department of Neurology The First Affiliated Hospital of Nanjing Medical University Nanjing Jiangsu 210029 China

Background： The functional improvement following bone marrow stromal cells （BMSCs） transplantation after stroke is directly related to the number of engrafted cells and neurogenesis in the injured brain. Here, we tried to evaluate whether 3-methyl-1-phenyl-2-pyrazolin-5-one （MCI-186）, a free radical scavenger, might influence BMSCs migration to ischemic brain, which could promote neurogenesis and thereby enhance treatment effects after stroke. Methods： Rat transient middle cerebral artery occlusion （MCAO） model was established. Two separate MCAO groups were administered with either MC1-186 or phosphate-buffered saline （PBS） solution to evaluate the expression of stromal cell-derived factor-1 （SDF-1） in ischemic brain, and compared to that in sham group （n 5/group/time point[at 1,3, and 7 days after operation]）. The content ofchemokine receptor-4 （CXCR4, a main receptor of SDF-I） at 7 days after operation was also observed on cultured BMSCs. Another four MCAO groups were intravenously administered with either PBS, MCI-186, BMSCs （2 ×106）, or a combination of MCI-186 and BMSCs （n = 10/ group）. 5-bromo-2-deoxyuridine （BrdU） and Nestin double-immunofluorescence staining was performed to identity the engrafted BMSCs and neuronal differentiation. Adhesive-removal test and foot-fault evaluation were used to test the neurological outcome. Results： MC1-186 upregulated the expression ofSDF- 1 in ischemic brain and CXCR4 content in BMSCs was enhanced after hypoxic stimulation. When MCAO rats were treated with either MCI- 186, BMSCs, or a combination ofMCI- 186 and BMSCs, the neurologic function was obviously recovered as compared to PBS control group （P 〈 0.01 or 0.05, respectively）. Combination therapy represented a further restoration, increased the number of BMSCs and Nestin- cells in ischemic brain as compared with BMSCs monotherapy （P 〈 0.01）. The number ofengrafted-BMSCs was correlated with the density of neuronal cells in ischemic brain （r = 0.72, P 〈 0.01 ） and the improvement of foot-thult （r = 0.70, P 〈 0.0 t ）. C

关键词： 3-Methyl-l-phenyl-2-pyrazolin-5-one bone marrow stromal cells Ischemic Stroke Neurogenesis stromal Cell-derivedFactor- 1

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Tissue Extracts From Infarcted Myocardium of Rats in Promoting the Differentiation of bone marrow stromal cells Into Cardiomyocyte-like cells

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Biomedical and Environmental Sciences 2008年第2期21卷 110-117页

作者： XIAO-NING LIU Oi YIN HAO ZHANG HONG ZHANG SHEN-JUN ZHU YING-Jie WEI SHENG-SHOU HU Key Laboratory of Cardiovascular Regenerative Medicine Ministry of Health and Department of Cardiovascular Surgery Cardiovascular Institute of Fuwai Hospital Chinese Academy of Medical Sciences ＆ Peking Union Medical College Beijing 100037 China Department of Thoracicand Cardiovascular Surgery FirstAffiliated Hospital Wenzhou Medical College Wenzhou 325000 Zhejiang China

Objective To investigate whether cardiac tissue extracts from rats could mimic the cardiac microenvironment and act as a natural inducer in promoting the differentiation of bone marrow stromal cells （BMSCs） into cardiomyocytes. Methods Three kinds of tissue extract or cell lysate [infarcted myocardial tissue extract （IMTE）, normal myocardial tissue extract （NMTE） and cultured neonatal myocardial lysate （NML）] were employed to induce BMSCs into cardiomyocyte-like cells. The cells were harvested at each time point for reverse transcription-polymerase chain reaction （RT-PCR） detection, immunocytochemical analysis, and transmission electron microscopy. Results After a 7-day induction, BMSCs were enlarged and polygonal in morphology. Myofilaments, striated sarcomeres, Z-lines, and more mitochondia were observed under transmission electron microscope. Elevated expression levels of cardiac-specific genes and proteins were also confirmed by RT-PCR and immunocytochemistry. Moreover, IMTE showed a greater capacity of differentiating BMSCs into cardiomyocyte-like cells. Conclusions Cardiac tissue extracts, especially IMTE, can effectively differentiate BMSCs into cardiomyocyte-like cells.

关键词： bone marrow stromal cells Cell differentiation Cardiac tissue extracts Myocardial infarction

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Study on the adoption of Schwann Cell Phenotype by bone marrow stromal cells in vitro and in vivo

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Biomedical and Environmental Sciences 2005年第5期18卷 326-333页

作者： FU-QIANG ZHAO PEI-XUN ZHANG XIANG-JUN HE CHAN DU ZHONG-GUO FU DIAN-YING ZHANG BAO-GUO JIANG People＇s Hospital Peking University Beijing 100044 China

To explore the possibilities of bone marrow stromal cells （MSCs） to adopt Schwann cell phenotype in vitro and in vivo in SD rats. Methods MSCs were obtained from tibia and femur bone marrow and cultured in culture flasks. Beta-mercaptoethanol followed by retinoic acid, forskolin, basic-FGF, PDGF and heregulin were added to induce differentiation of MSCs＇. Schwann cell markers, p75, S-100 and GFAP were used to discriminate induced properties of MSCs＇ by immunofluorescent staining. PKH-67-1abelled MSCs were transplanted into the mechanically injured rat sciatic nerve, and laser confocal microscopy was performed to localize the PKH67 labelled MSCs in the injured sciatic nerve two weeks after the operation. Fluorescence PKH67 attenuation rule was evaluated by flow cytometry in vitro. Results MSCs changed morphologically into cells resembling primary cultured Schwann cells after their induction in vitro. In vivo, a large number of MSCs were cumulated within the layer of epineurium around the injured nerve and expressed Schwann cell markers, p75, S- 100, and GFAP. Conclusion MSCs are able to support nerve fiber regeneration and re-myelination by taking on Schwann cell function, and can be potentially used as possible substitutable cells for artificial nerve conduits to promote nerve regeneration.

关键词： bone marrow stromal cells Differentiation Schwann cells

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Granulocyte-macrophage colony-stimulating factor-transfected bone marrow stromal cells for the treatment of ischemic stroke

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Neural Regeneration Research 2012年第16期7卷 1220-1227页

作者： Xingjian Lin Yingdong Zhang Weiguo Liu Jingde Dong Jie Lu Qing Di Jingping Shi Nanjing Medical University Affiliated Nanjing Brain Hospital Department of Neurology Nanjing 210029 Jiangsu Province China Nanjing Medical University Affiliated Nanjing First Hospital Department of Neurology Nanjing 210029 Jiangsu Province China

Adult, male, Sprague-Dawley rats were injected with granulocyte-macrophage colony-stimulating factor-transfected bone marrow stromal cells （GM-CSF-BMSCs） into the ischemic boundary zone at 24 hours after onset of middle cerebral artery occlusion. Results showed reduced infarct volume, decreased number of apoptotic cells, improved neurological functions, increased angiogenic factor expression, and increased vascular density in the ischemic boundary zone in rats that underwent GM-CSF-BMSCs transplantation compared with the BMSCs group. Experimental findings suggested that GM-CSF-BMSCs could serve as a potential therapeutic strategy for ischemic stroke and are superior to BMSCs alone.

关键词： bone marrow stromal cells granulocyte-macrophage colony-stimulating factor gene transfection ischemic stroke transplantation stem cells neural regeneration

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Transfect bone marrow stromal cells with pcDNA3.1-VEGF to construct tissue engineered bone in defect repair

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Chinese Medical Journal 2012年第5期125卷 906-911页

作者： SI Hai-peng LU Zhi-hua LIN Yong-liang LI Jing-jing YIN Qing-feng ZHAO Dong-mei WANG Shao-jin LI Jian-min WANG Hai-bin ZHANG Xi-hua Department of Orthopedics the Second Hospital of ShandongUniversity Jinan Shandong 250033 China Department of Physics and Information Engineering JiningUniversity Jining Shandong 272009 China Medical School of Shandong University Jinan shandong 250012China Qilu Hospital of Shandong University Jinan Shandong 250012China Materials Science and Engineering School of Shandong University Jinan Shandong 250061 China

Background We previously showed that nano-hydroxyapatite/carboxymethyl chitosan （n-Ha/CMCS） displayed excellent mechanical properties, good degradation rates and exceptional biocompatibility, with negligible toxicity. The aim of this study was to determine the effect of the same composite with vascular endothelial growth factor （VEGF）transfected bone marrow stromal cells （BMSCs） in a rabbit radial defect model.

关键词： bone defect bone marrow stromal cells plasmid bone tissue engineering repair genipin

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Direct contact with bone marrow stromal cells promotes the invasions of SHI-1 leukemia cells

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Chinese Medical Journal 2013年第14期126卷 2731-2735页

作者： LI Zhen-jiang CHEN Zi-xing CEN Jian-nong HE Jun QIU Qiao-cheng Department of Hematology Second Affiliated Hospital of Nanchang University Nanchang Jiangxi 330006 China Leukemia Research Unit Jiangsu Institute of Hematology First Affiliated Hospital of Soochow University Suzhou Jiangsu 215006 China

Background Interactions of tumor cells with the microenvironment were deemed to promote the tumor invasion and metastasis. CXC chemokine receptor 4 （CXCR4） and extracellular matrix metalloproteinase inducer （EMMPRIN） had reported to participate in this process. However the roles of bone marrow microenvironment in leukemic infiltration were not well investigated. Methods A co-culture system between SHI-1 cells and bone marrow stromal cells （BMSCs） is used to simulate the interactions of leukemic cells with their microenvironment. The trans-matrigel invasion was used to detect the capability of SHI-1 cells invasion. The BMSCs and SHI-1 cells were mixed in a ratio of 1：10 and added to the millicell chamber coated with matrigel. Either the co-culture supernatant or the functional blocking peptide of CXCR4 and EMMPRIN were added to the trans-matrigel invasion system. The expressions of EMMPRIN in SHI-1 cells and BMSCs were detected by RT-PCR. The changes of the expression of matrix metalloproteinase-2, 9 （MMP-2, MMP-9）, tissue inhibitor of metalloproteinase 2 （TIMP-2）, and CXCR4 mRNA in SHI-1 cells were determined by real-time PCR. The concentration of stromal cell derived factor 1 （SDF-1） in serum free supernatant was measured by ELISA. Results Both SHI-1 cells and BMSCs express EMMPRIN. SHI-1 cells could hardly invade the matrigel membrane; the co- culture supernatant did not induce the invasion of SHI-1 cells. When contacting directly with BMSCs, SHI-1 cells invaded to the lower chamber of millicell were significantly increased. The functional blocking peptide of CXCR4 and EMMPRIN could significantly inhibit the invasion triggered by BMSCs. When co-culturing with BMSCs, the expression of CXCR4, MMP-2, MMP-9 and TIMP-2 mRNA in SHI-1 cells were significantly elevated in company with a significantly higher level of SDF-1 in the co-cultured serum-free supernatant. Conclusion The interactions of leukemic cells and BMSCs play important roles in leukemic cell infiltration.

关键词： leukemia bone marrow stromal cells extramedullary infiltration EMMPRIN SDF-1/CXCR4

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Effects of endothelin-1 on differentiation of cardiac myocyte induced from rabbit bone marrow stromal cells

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Chinese Medical Journal 2006年第10期119卷 832-839页

作者： DONG Hong-yan ZHANG Zhong-ming ZHOU Zhong-xin Centre of Neurobiological Research Xuzhou Medical CollegeXuzhou 221002 China Department of Thoracic Cardiovascular Surgery AffiliatedHospital of Xuzhou Medical College Xuzhou 221002 China

Background Cardiomyocyte transplantation for the therapy of myocardial ischaemia is being paid close attention. However, how the microenvironment controls the differentiation of transplanted bone marrow stromal cells （BMSCs） is unknown. Endothelin-1 （ET-1）, a cytokine, increases during myocardial infarction, but it is not known whether ET-1 is responsible for the fate of transplanted BMSCs. In the present study, we investigated the effects of ET-1 on differentiation and maturation of induced rabbit BMSCs, in vitro, to elucidate the cellular biological mechanisms.

关键词： bone marrow stromal cells endothelin-1 differentiation cell culture

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