scholarly journals Marrow-Derived Mesenchymal Stem Cells Transfected with Survinvin Gene Protect Kidney from ischemia-reperfusion Injury in Rats

2021 ◽  
Author(s):  
Yang Xiaofeng ◽  
Qianqian Wang
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Renal Tubule ◽  
Ischemia Reperfusion Injury ◽  
Lentiviral Vector ◽  
Ischemia Reperfusion ◽  
Renal Ischemia ◽  
The Self ◽  
The Other ◽  
Control Group

Abstract Objective: To investigate the survival ability of bone marrow Mesenchymal stem cells (MSCs) transfected with survinvin gene in the microenvironment of renal ischemia,and to study the ability and mechanism of repairing renal ischemia-reperfusion injury in rats.Method: Mesenchymal stem cells (MSCs) from bone marrow of male Sprague–Dawley rat were infected with the self‐inactive lentiviral vector and transfected with the Survinvin gene recombinant vector and then EGFP-tagged. After amplification and culture, they were detected by green fluorescence and then retained.48 specific pathogen-free C57BL/6J mice were randomly divided into 4 groups of 12 each. Rats in the control group were only surgically exposed. The other 3 groups were surgically exposed and the bilateral renal arteries were clamped for 45 minutes to restore blood supply, and models of renal ischemia-reperfusion were established. There were control group,ischemia reperfusion group(Marked as IR group), empty virus transfection transplantation group(Marked as MSCs group) or survinvin gene transfection transplantation group(Marked as SVV/MSCs group), and sequentially injected with normal saline,normal saline,1×106 MSCs infected with the self‐inactive lentiviral vector or 1×106 survivin gene-expressing MSCs. At different time points of 1d, 3d, 7d, 14d, collect serum to test blood urea nitrogen detection, to cut the rat kidney section for quantitative analysis, HE staining to observe renal issues changes and the degree of renal tubular damage and IL-10 by using ELISA detection. Result: The MSCs with resuscitation and expansion culture had strong proliferation and good fluorescence. The creatinine urea nitrogen level in the MSCs group and SVV/MSCs group was significantly lower than that in the IR group and control group (p<0.01 or p<0.05). The pathological damage score of HE staining in the kidney was lighter in the stem cell transplantation group, and the SVV/MSCs group was significantly lower than the other two groups (p<0.01 or p<0.05). On the 3rd and 14th day, the number of transplanted cells in the kidney tissue was much higher in the SVV/MSCs group than in the MSCs group. The MSCs expressing EGFP were mainly distributed around the glomerulus, the small vessel inner wall, and the interstitial between the renal tubule and the renal tubule. However, MSCs expressing EGFP were hardly seen on the inner wall of the renal tubule. The levels of protective factors IL-10 increased after renal ischemic injury. SVV/MSCs group was also significantly more than IR group or MSCs group (P<0.01 or p<0.05). And there was no statistical difference from the normal control group on the 14th day.Conclusion: Transfection of Survinifin gene can increase the survival ability of MSCs in ischemic kidney. After transplantation, MSCs are not directly differentiated into injured tubular endothelial cells, which further promote the repair of kidney damage through its strong paracrine effect.

scholarly journals Effect of Lentivirus-mediated GDF5 Transfection on Differentiation of Rabbit Nucleus Pulposus Mesenchymal Stem Cells

2020 ◽  
Author(s):  
kun zhu ◽  
Rui Zhao ◽  
Yuchen Ye ◽  
Gang Xu ◽  
Changchun Zhang
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Nucleus Pulposus ◽  
Lentiviral Vector ◽  
Intervertebral Discs ◽  
Degenerative Disease ◽  
The Other ◽  
Control Group ◽  
Nucleus Pulposus Cells ◽  
Qrt Pcr

Abstract Background: Disc degenerative disease is a common senile degenerative disease, which seriously affects the quality of life of patients.The purpose of this study is to observe the biological and cytological characteristics of rabbit nucleus pulposus mesenchymal stem cells (NPMSCs), and to determine the effect of growth differentiation factor 5(GDF5) on the differentiation of rabbit NPMSCs by lentivirus transfection.Methods: In vitro culture model of rabbit NPMSCs was established and NPMSCs cells were identified by flow cytometry (FCM)and quantitative real-time PCR(qRT-PCR). Then NPMSCs were divided into three groups: lentiviral vector carrying GDF5 was used to transfect NPMSCs, to determine the transfection rate, which was recorded as transfection group, and the NPMSCs transfected with ordinary lentiviral vector was recorded as control group, NPMSCs without processing was recorded as normal group. FCM, qRT-PCR and Western Blot(WB) were used to detected the change of NPMSCs.Results: The transfected NPMSCs by GDF5 became longer and narrower, and the cell density decreased,and the positive rate of GDF5 in the transfected group was significantly higher than that in the other two groups (P<0.05). The mRNA expression of KRT8, KRT18, KRT19 in the transfected group was significantly higher than the other two groups(P<0.05),the result of WB were the same to qRT-PCR. Conclusions: GDF5 can induce the differentiation of NPMSCs and repair degenerative intervertebral discs. Lentiviral vector carrying GDF5 can be integrated into the chromosome genome of NPMSCs and promote differentiation of NPMSCs into nucleus pulposus cells(NPCs).

scholarly journals Intravenously delivered mesenchymal stem cells prevent MVO formation after myocardial ischemia/reperfusion injury

2020 ◽  
Vol 41 (Supplement_2) ◽  
Author(s):  
J Xie
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Reperfusion Injury ◽  
Nk Cells ◽  
Ischemia Reperfusion Injury ◽  
Ischemia Reperfusion ◽  
Group Versus ◽  
Control Group ◽  
Funding Source ◽  
Ischemia And Reperfusion Injury

Abstract   Microvascular obstruction (MVO) after primary PCI is identified as an independent risk factor for prognosis in AMI patients. Inflammatory response induced by ischemia and reperfusion injury (I/R injury) was considered as one of main mechanisms for MVO formation. Mesenchymal stem cells (MSCs) are a unique stromal cell type that confers immunomodulatory effect in cardiac disease. Here we investigate whether intravenously and immediately delivered MSC could be used as potential therapeutic method to attenuate MVO formation. A cardiac-catheterization-induced porcine model of myocardial I/R injury was established, and allograft MSCs were delivered intravenously and immediately. Cardiac magnetic resonance imaging was performed 2 days and 7 days after operation to determine infarct area, MVO and cardiac function. We observed that the animals with allograft MSC delivering revealed decreased MVO and infarct size, as well as improved LVEF. In histology analysis, decreased myocytes area, fibrosis and inflammatory cell infiltration were observed in peri-infarct zone of animals with allograft MSC delivering. Meanwhile, the concentrations of IL-6, CRP and IL-1β in the serum were reduced in the allograft MSC group versus the control group. Flow cytometry indicated that decreased NK cells in the peripheral blood and ischemic heart tissue in the animals with allograft MSC delivering. In summary, we observed that allograft MSC delivering intravenously and immediately after myocardium I/R injury could attenuate MVO formation in porcine by NK cells depression. Figure 1 Funding Acknowledgement Type of funding source: Public grant(s) – National budget only. Main funding source(s): Natural Science Foundation of China

Modulation of renal ischemia/reperfusion in rats by a combination of ischemic preconditioning and adipose-derived mesenchymal stem cells (ADMSCs)

2016 ◽  
Vol 94 (9) ◽  
pp. 936-946 ◽  
Author(s):  
Abdelaziz M. Hussein ◽  
Nashwa Barakat ◽  
Amira Awadalla ◽  
Mahmoud M. Gabr ◽  
Sherry Khater ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Ischemic Preconditioning ◽  
Caspase 3 ◽  
Ischemia Reperfusion ◽  
Treated Group ◽  
Renal Ischemia ◽  
Control Group ◽  
Sprague Dawley ◽  
Sprague Dawley Rats

The present study investigated the effects of combination of ischemic preconditioning (Ipre) and adipose-derived mesenchymal stem cells (ADMSCs) on renal ischemia–reperfusion (I–R) injury in rats. 90 male Sprague Dawley rats were divided into 5 equal groups; sham operated, control (45 min left renal ischemia), Ipre group as control group with 3 cycles of Ipre just before renal ischemia, ADMSCs-treated group (as control with ADMSCs 106 cells in 0.1 mL via penile vein 60 min before ischemia time), and Ipre + ADMSCs group as ADMCs group with 3 cycles of Ipre. Ipre and ADMSCs groups showed significant decrease in serum creatinine and blood urea nitrogen (BUN) and caspase-3 and CD45 expression in kidney and significant increase in HIF-1α, SDF-1α, CD31, and Ki67 expressions in kidney compared with the control group (p < 0.05). Moreover, the Ipre + ADMSCs group showed significant decrease in serum BUN and caspase-3 and CD45 expression in kidney with significant increase in HIF-1α, SDF-1α, CD31, and Ki67 expression in kidney compared with the Ipre and ADMCs groups (p < 0.05). We concluded that Ipre potentiates the renoprotective effect of ADMSCs against renal I/R injury probably by upregulation of HIF-1α, SDF-1α, CD31, and Ki67 and downregulation of caspase-3 and CD45.

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