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Hydrogen peroxide preconditioning enhances the therapeutic efficacy of Wharton＇s Jelly mesenchymal stem cells after myocardial infarction

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Chinese Medical Journal 2012年第19期125卷 3472-3478页

作者： ZHANG Jin CHEN Guang-hui WANG Yong-wei ZHAO Jing DUAN Hai-feng LIAO Lian-ming ZHANG Xiao-zhong CHEN Yun-dai CHEN Hu Department of Cardiology Chinese People's Liberation Army General Hospital Beijing 100853 China Department of Emergency Chinese People's Liberation Army General Hospital Beijing 100853 China Department of Cardiology Affiliated Hospital Academy of Military Medical Sciences Beijing 100071 China Department of Hematopoietic stem Cell Transplantation Affiliated Hospital Academy of Military Medical Sciences Beijing 100071 China Department of Experimental Hematology Institute of Radiation Medical Sciences Academy of Military Medical Sciences Beijing 100850 China Institute of Oncology Academy of Integrative Medicine Fujian University of Traditional Chinese Medicine Fuzhou Fujian 350122 China

Background Exposure of cells to sublethal concentrations of hydrogen peroxide （H202） can alleviate subsequent oxidative stress-induced apoptosis. We assessed the effects of H202 preconditioning on the therapeutic potential of human umbilical cord Wharton＇s Jelly mesenchymal stem cells （WJ-MSCs） in a murine model of myocardial infarction. Methods WJ-MSCs were incubated in the media for 2 hours with or without 200 iJmol/L H202. Mice underwent left anterior descending coronary artery ligation, and received injection of phosphate buffered saline, 1×10^6 WJ-MSCs, or 1×10^6 H202 preconditioned WJ-MSCs 3 hours later via tail vein. Echocardiography was performed 0, 7, 14 and 28 days after surgery, and the mice were euthanized on day 28 for histological analysis. In vitro cytokine concentrations in the WJ-MSC cell supernatant were measured by enzyme-linked immunosorbent assay （ELISA）. The effect of WJ-MSC cell supernatant on the migration and proliferation of endothelial cells were observed by transwell migration and 3-（4, 5-dimethylthiazol-2-yl）-2, 5-diphenyl tetrazoliumbromide （MTT） assays. Results Echocardiographic measurements revealed a significant improvement in the left ventricular contractility of the WJ-MSCs-H2O2 group compared to the WJ-MSCs group. Histological analysis revealed increased neovascularization and reduced myocardial fibrosis in the WJ-MSCs-H2O2 group compared to the WJ-MSCs group. Pretreatment of WJ-MSCs with H2O2 increased the secretion of interleukin-6 （IL-6） into the cell culture supernatant by approximately 25-fold. The culture supernatant from WJ-MSCs-H2O2 significantly increased the migration and proliferation of endothelial cells; these effects could be blocked using an anti-IL-6 antibody. Conclusions This study demonstrates that H2O2 preconditioning significantly enhanced the therapeutic potential of WJ-MSCs, possibly by stimulating the production of IL-6 by WJ-MSCs, which may cause migration and proliferation of endothelial cells and increase neovascularization.

关键词： mesenchymal stem cells reactive oxygen species preconditioning angiogenesis

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Injection of bone marrow mesenchymal stem cells by intravenous or intraperitoneal routes is a viable alternative to spinal cord injury treatment in mice

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Neural Regeneration Research 2018年第6期13卷 1046-1053页

作者： Bruna dos Santos Ramalho FernANDa Martins de Almeida Conrado Mendonca Sales Silmara de Lima Ana Maria Blanco Martinez Laboratorio de Neurodegeneracao e Reparo Departamento de Patologia - Faculdade de Medicina HUCFF Universidade Federal do Rio de Janeiro Rio de laneiro Brazil

In spite of advances in surgical care and rehabilitation, the consequences of spinal cord injury （SCI） are still challenging. Several experimental therapeutic strategies have been studied in the SCI field, and recent advances have led to the development of therapies that may act on the inhibitory microenvironment. Assorted lineages of stem cells are considered a good treatment for SCI. This study investigated the effect of systemic transplantation of mesenchymal stem cells （MSCs） in a compressive SCI model. Here we present results of the intraperitoneal route, which has not been used previously for MSC administration after compressive SCI. We used adult female C57BL/6 mice that underwent laminectomy at the T9 level, followed by spinal cord compression for 1 minute with a 30-g vascular clip. The animals were divided into five groups： sham （anesthesia and laminectomy but without compression injury induction）, MSC i.p. （intraperitoneal injection of 8×10^5 MSCs in 500 μL of DMEM at 7 days after SCI）, MSC i.v. （intravenous injection of 8 × 10^5 MSCs in 500μL of DMEM at 7 days after SCI）, DMEM i.p. （intraperitoneal injection of 500μL of DMEM at 7 days after SCI）, DMEM i.v. （intravenous injection of 500 μL of DMEM at 7 days after SCI）. The effects of MSCs transplantation in white matter sparing were analyzed by luxol fast blue staining. The number of preserved fibers was counted in semithin sections stained with toluidine blue and the presence of trophic factors was analyzed by immunohistochemistry. In addition, we analyzed the locomotor performance with Basso Mouse Scale and Global Mobility Test. Our results showed white matter preservation and a larger number of preserved fibers in the MSC groups than in the DMEM groups. Furthermore, the MSC groups had higher levels of trophic factors （brain-derived neurotrophic factor, nerve growth factor, neurotrophin-3 and neurotrophin-4） in the spinal cord and improved locomotor performance. Our results indicate that injection of MSCs by either intraperitoneal or intravenous route

关键词： spinal cord injury mesenchymal stem cells intravenous route intraperitoneal route functional recovery systemic transplantation neural regeneration

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Bone marrow-derived mesenchymal stem cells migrate to healthy and damaged salivary glands following stem cell infusion

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International Journal of Oral Science 2014年第3期6卷 154-161页

作者： Silke Schwarz Ralf Huss Michaela Schulz-Siegmund Breda Vogel Sven Brandau Stephan Lang Nicole Rotter Department of Otorhinolaryngology Ulm University Medical Center Apceth Munich Germany Institute of Pharmaceutical Technology University of Leipzig Department of Otorhinolaryngology University of Duisburg–Essen

Xerostomia is a severe side effect of radiation therapy in head and neck cancer patients. To date, no satisfactory treatment option has been established. Because mesenchymal stem cells（MSCs） have been identified as a potential treatment modality, we aimed to evaluate stem cell distribution following intravenous and intraglandular injections using a surgical model of salivary gland damage and to analyse the effects of MSC injections on the recruitment of immune cells. The submandibular gland ducts of rats were surgically ligated. Syngeneic adult MSCs were isolated, immortalised by simian virus 40（SV40） large T antigen and characterized by flow cytometry. MSCs were injected intravenously and intraglandularly. After 1, 3 and 7 days, the organs of interest were analysed for stem cell recruitment. Inflammation was analysed by immunohistochemical staining. We were able to demonstrate that, after intravenous injection, MSCs were recruited to normal and damaged submandibular glands on days 1, 3 and 7. Unexpectedly, stem cells were recruited to ligated and non-ligated glands in a comparable manner. After intraglandular injection of MSCs into ligated glands, the presence of MSCs, leucocytes and macrophages was enhanced, compared to intravenous injection of stem cells. Our data suggest that injected MSCs were retained within the inflamed glands, could become activated and subsequently recruited leucocytes to the sites of tissue damage.

关键词： duct ligation mesenchymal stem cells submandibular gland injury xerostomia

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Detection of Repair of the Zone of Calcified Cartilage with Osteoarthritis through mesenchymal stem cells by Ultrashort Echo Time Magnetic Resonance Imaging

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Chinese Medical Journal 2018年第9期131卷 1092-1098页

作者： Quan Zhou Shao-Lin Li Ya-Jun Ma Vicki de Tal Wei Li Ying-Hua Zhao Department of Radiology Third Affiliated Hospital of Southern Medical University (Academy of Orthopedics Guangdong Province) Guangzhou Guangdong 510630 China Department of Medical Imaging The Fifth Affiliated Hospital of Sun Yat-sen University Zhuhai Guangdong 519000 China Department of Radiology University of California San Diego CA 92103-8226 USA

Objective：Currently,magnetic resonance imaging （MRI） is the most commonly used imaging modality for observing the growth and development of mesenchymal stem cells （MSCs） after in vivo transplantation to treat osteoarthritis （OA）.However,it is a challenge to accurately monitor the treatment effects of MSCs in the zone of calcified cartilage （ZCC） with OA.This is especially true in the physiological and biochemical views that are not accurately detected by MRI contrast agents.In contrast,ultrashort time echo （UTE） MRI has been shown to be sensitive to the presence of the ZCC,creating the potential for more effectively observing the repair of the ZCC in OA by MSCs.A special focus is given to the outlook of the use ofUTE MRI to detect repair of the ZCC with OA through MSCs.The limitations of the current techniques for clinical applications and future directions are also discussed.Data Sources：Using the combined keywords：＂osteoarthritis＂,＂mesenchymal stem cells＂,＂calcified cartilage＂,and ＂magnetic resonance imaging＂,the PubMed/MEDLINE literature search was conducted up to June 1,2017.Study Selection：A total of 132 published articles were initially identified citations.Of the 132 articles,48 articles were selected after further detailed review.This study referred to all the important English literature in full.Results：In contrast,UTE MRI has been shown to be sensitive to the presence of the ZCC,creating the potential for more effectively observing the repair of the ZCC in OA by MSCs.Conclusions：The current studies showed that the ZCC could be described in terms of its histomorphology and biochemistry by UTE MRI.We prospected that UTE MRI has been shown the potential for more effectively observing the repair of the ZCC in OA by MSCs in vivo.

关键词： Magnetic Resonance Imaging mesenchymal stem cells Osteoarthritis Ultrashort Echo Time Zone of Calcified Cartilage

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Human umbilical cord mesenchymal stem cells and the treatment of spinal cord injury

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Chinese Medical Journal 2009年第2期122卷 225-231页

作者： CAO Fu-jiang FENG Shi-qing Department of Orthopaedics General Hospital of Tianjin Medical University Tianjin 300052 China

Objective To review the recent studies about human umbilical cord mesenchymal stem cells （hUCMSCs） and advances in the treatment of spinal cord injury. Data sources Published articles （1983-2007） about hUCMSCs and spinal cord injury were selected using Medline. Study selection Articles selected were relevant to development of mesenchymal stem cells （MSCs） for transplantation in spinal cord injury therapy. Of 258 originally identified articles 51 were selected that specifically addressed the stated purpose. Results Recent work has revealed that hUCMSCs share most of the characteristics with MSCs derived from bone marrow and are more appropriate to transplantation for cell based therapies. Conclusions Human umbilical cord could be regarded as a source of MSCs for experimental and clinical needs. In addition, as a peculiar source of stem cells, hUCMSCs may play an important role in the treatment of spinal cord injury. Chin Med J 2009;122（2）：225-231

关键词： umbilical cord mesenchymal stem cells spinal cord injuries

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The role of mitochondria in osteogenic, adipogenic and chondrogenic differentiation of mesenchymal stem cells

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Protein & Cell 2017年第6期8卷 439-445页

作者： Qianqian Li Zewen Gao Ye Chen Min-Xin Guan Division of Clinical Genetics and Genomics The Children's Hospital Zhejiang University School of MedicineHangzhou 310058 China Institute of Genetics Zhejiang University and Department of Genetics Zhejiang University School of MedicineHangzhou 310058 China

mesenchymal stem cells （MSCs） are progenitors of con- nective tissues, which have emerged as important tools for tissue engineering due to their differentiation potential along various cell types. In recent years, accumulating evidence has suggested that the regulation of mitochon- dria dynamics and function is essential for successful dif- ferentiation of MSCs. In this paper, we review and provide an integrated view on the role of mitochondria in MSC dif- ferentiation. The mitochondria are maintained at a relatively low activity level in MSCs, and upon induction, mtDNA copy number, protein levels of respiratory enzymes, the oxygen consumption rate, mRNA levels of mitochondrial biogene- sis-associated genes, and intracellular ATP content are increased. The regulated level of mitochondrial ROS is found not only to influence differentiation but also to con- tribute to the direction determination of differentiation. Understanding the roles of mitochondrial dynamics during MSC differentiation will facilitate the optimization of differ- entiation protocols by adjusting biochemical properties, such as energy production or the redox status of stem cells, and ultimately, benefit the development of new pharmacologic strategies in regenerative medicine.

关键词： mesenchymal stem cells mitochondria,differentiation

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Notch signaling： a novel regulating differentiation mechanism of human umbilical cord blood-derived mesenchymal stem cells into insulin-producing cells in vitro

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Chinese Medical Journal 2010年第5期123卷 606-614页

作者： HU Yan-hua WU De-quan GAO Feng LI Guo-dong ZHANG Xin-chen Department of General Surgery Second Affiliated Hospital of Harbin Medical University Harbin Heilongjiang 150086 China

Background Human umbilical cord blood-derived mesenchymal stem cells （UCB-MSCs） could be induced to differentiate into insulin producing cells （IPCs） in vitro, which have good application potential in the cell replacement treatment of type-1 diabetes. However, the mechanisms regulating this differentiation have remained largely unknown. Notch signaling is critical in cell differentiation. This study investigated whether Notch signaling could regulate the IPCs differentiation of human UCB-MSCs. Methods Using an interfering Notch signaling protocol in vitro, we studied the role of Notch signaling in differentiation of human UCB-MSCs into IPCs. In a control group the induction took place without interfering Notch signaling. Results Human UCB-MSCs expressed the genes of Notch receptors （Notch 1 and Notch 2） and ligands （Jagged 1 and Deltalike 1）. Human UCB-MSCs with over-expressing Notch signaling in differentiation resulted in the down-regulation of insulin gene level, proinsulin protein expression, and insulin-positive cells percentage compared with the control group. These results showed that over-expressing Notch signaling inhibited IPCs differentiation. Conversely, when Notch signaling was attenuated by receptor inhibitor, the induced cells increased on average by 3.06-fold （n=-4, P 〈0.001） in insulin gene level, 2.60-fold （n=-3, P 〈0.02） in proinsulin protein expression, and 1.62-fold （n=-6, P 〈0.001） in the rate of IPCs compared with the control group. Notch signaling inhibition significantly promoted IPCs differentiation with about 40% of human UCB-MSCs that converted to IPCs, but these IPCs were not responsive to glucose challenge very well both in vitro and in vivo. Hence, further research has to be carried out in the future. Conclusions Notch signaling may be an important mechanism regulating IPCs differentiation of human LICB-MSCs in vitro and Notch signaling inhibition may be an efficient way to increase the number of IPCs, which may resolve the shortage of islet of cell replacement treatment of type-1 d

关键词： Notch signaling mesenchymal stem cells differentiation insulin producing cells diabetes

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Differentiation of bone marrow-derived mesenchymal stem cells from diabetic patients into insulin-producing cells in vitro

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Chinese Medical Journal 2007年第9期120卷 771-776页

作者： SUN Yu CHEN Li HOU Xin-guo HOU Wei-kai DONG Jian-jun SUN Lei TANG Kuan-xiao WANG Bin SONG Jun LI Hui WANG Ke-xin Department of Endocrinology Qilu Hospital Shandong University Jinan 250012 China Yuhuangding Hospital Yantai 264000 China

Bckground stem cells, which have the ability to differentiate into insulin-producing cells （IPCs）, would provide a potentially unlimited source of islet cells for transplantation and alleviate the major limitations of availability and allogeneic rejection. Therefore, the utilization of stem cells is becoming the most promising therapy for diabetes mellitus （DM）. Here, we studied the differentiation capacity of the diabetic patient＇s bone marrow-derived mesenchymal stem cells （MSCs） and tested the feasibility of using MSCs for β-cell replacement. Methods Bone marrow-derived MSCs were obtained from 10 DM patients （5 type 1 DM and 5 type 2 DM） and induced to IPCs under a three-stage protocol. Representative cell surface antigen expression profiles of MSCs were analysed by flow cytometric analysis. Reverse transcription-polymerase chain reaction （RT-PCR） was performed to detect multiple genes related to pancreatic β-cell development and function. The identity of the IPCs was illustrated by the analysis of morphology, ditizone staining and immunocytochemistry. Release of insulin by these cells was confirmed by immunoradioassay. Results Flow cytometric analysis of MSCs at passage 3 showed that these cells expressed high levels of CD29 （98.28%）, CD44 （99.56%） and CD106 （98.34%）. Typical islet-like cell clusters were observed at the end of the protocol （18 days）. Ditizone staining and immunohistochemistry for insulin were both positive. These differentiated cells at stage 2 （10 days） expressed nestin, pancreatic duodenal homeobox-1 （PDX-1）, Neurogenin3, Pax4, insulin, glucagon, but at stage 3 （18 days） we observed the high expression of PDX-1, insulin, glucagon. Insulin was secreted by these cells in response to different concentrations of glucose stimulation in a regulated manner （P〈0.05）. Conclusions Bone marrow-derived MSCs from DM patients can differentiate into functional IPCs under certain conditions in vitro. Using diabetic patient＇s own bone marrow-derived MSCs as a source of autologous IPCs for β-cell reDl

关键词： diabetes mellitus mesenchymal stem cells insulin transdifferentiation

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Short-term memory of danger signals or environmental stimuli in mesenchymal stem cells： implications for therapeutic potential

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Cellular & Molecular Immunology 2016年第3期13卷 369-378页

作者： Guang-Yang Liu Yang Liu Ying Lu Ya-Ru Qin Guo-Hu Di Yong-Hong Lei Hu-Xian Liu Yan-Qi Li Chutse Wu Xian-Wen Hu Hai-Feng Duan Key Laboratory of Systems Bioengineering School of Chemical Engineering and Technology Tianjin University Tianjin China Beijing Institute of Radiation Medicine (BIRM) Beijing China 307 Hospital of the Chinese People's Liberation Army Beijing China 301 Hospital of the Chinese People's Liberation Army Beijing China Beijing Institute of Biotechnology (BIB) Beijing China

mesenchymal stem/stromal cells （MSCs） possess some characteristics of immune cells, including a pro-inflammatory phenotype, an immunosuppressive phenotype, antibacterial properties and the expression of Toll-like receptor proteins. Here we show that, similar to immune cells, MSCs retain information from danger signals or environmental stimuli for a period of time. When treated with the pro-inflammatory factors lipopolysaccharide （LPS） or tumor necrosis factor-a （TNF-a）, MSCs display increased expression of IL-6, IL-8 and MCP-1. Following re-plating and several rounds of cell division in the absence of stimulating factors, the expression of IL-6, IL-8 and MCP-1 remained higher than in untreated cells for over 7 days. A spike in cytokine secretion occurred when cells were exposed to a second round of stimulation. We primed MSCs with LPS and LPS-primed MSCs had better therapeutic efficacy at promoting skin flap survival in a diabetic rat model than did unprimed MSCs. Finally, we found that several microRNAs, including miR146a, miR150 and miR155, along with the modification of DNA by 5-hydroxymethylcytosine （5hmC）, mediate the MSC response to LPS and TNF-e stimulation. Collectively, our data suggest that MSCs have a short-term memory of environmental signals, which may impact their therapeutic potential.

关键词： environmental stimuli mesenchymal stem cells pre-conditioning short-term memory

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Dickkopf-1 has an Inhibitory Effect on mesenchymal stem cells to Fibroblast Differentiation

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Chinese Medical Journal 2016年第10期129卷 1200-1207页

作者： Yan Li Sang-Sang Qiu Yan Shao Hong-Huan Song Gu-Li Li Wei Lu Li-Mei Zhu Department of Chronic Communicable Disease Jianqsu Provincial Center for Disease Prevention and Control Nanjing Jiangsu 210009 China Department of Infection Management Affiliated Wuxi People's Hospital to Nanjing Medical University Wuxi Jiangsu 214023 China

Background： mesenchymal stem cells （MSCs） are bone marrow stem cells which play an important role in tissue repair. The treatment with MSCs will be likely to aggravate the degree of fibrosis. The Wnt/β-catenin signaling pathway is involved in developmental and physiological processes, such as fibrosis. Dickkopfs （DKKs） are considered as an antagonist to block Wnt/β-catenin signaling pathway by binding the receptor of receptor-related protein （LRP5/6）. DKK1 was chosen in attempt to inhibit fibrosis of MSCs by lowering activity of Wnt/β-catenin signaling pathway. Methods： Stable MSCs were randomly divided into four groups： MSCs control, MSCs ＋ transforming growth factor-β （TGF-β）, MSCs ＋ DKK1, and MSCs ＋ TGF-β ＋ DKK1. Flow cytometry was used to identify MSCs. Cell viability was evaluated by 3-（4,5-dimethyl-2-thiazolyl）-2,5-diphenyl-2-H-tetrazolium bromide test. lmmunofluorescence was used to detect protein expression in the Wnt/β-catenin signaling pathways. Western blotting analysis was employed to test expression of fibroblast surface markers and, finally, real-time reverse transcription polymerase chain reaction was employed to test mRNA expression offibroblast surface markers and Wnt/β-catenin signaling proteins. Results： Cultivated MSCs were found to conform to the characteristics of standard MSCs： expression of cluster of differentiation （CD） 73, 90, and 105, not expression of 34, 45, and 79. We found that DKK1 could maintain the normal cell morphology of MSCs. Western blotting analysis showed that fibroblast surface markers were expressed in high quantities in the group MSCs ＋ TGF-β. However, the expression was lower in the MSCs ＋ TGF-β ＋ DKK1. Immunofluorescence showed high expression of all Wnt/β-catnin molecules in the MSCs ＋ TGF-β group but expressed in lower quantities in MSCs ＋ TGF-β ＋ DKK1 group. Finally, mRNA expression of fibroblast markers vimentin, α-smooth muscle actin and Wnt/β-catenin signaling proteins β-catenin, T-cell factor, and glycogen synthase kinase-3β was significantly increased in MSCs

关键词： Dickkopfs-1 Fibrosis mesenchymal stem cells Wnt/β-catenin Signaling Pathway

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