Journal: Neurologia medico-chirurgica

Article Title: Recent Progress in Endothelial Progenitor Cell Culture Systems: Potential for Stroke Therapy

doi: 10.2176/nmc.ra.2016-0027

Figure Lengend Snippet: Chronological development of EPC-based cell therapies. Since EPCs were first isolated from peripheral blood, various EPC-based cell therapies have been developed. The latest fourth-generation EPC therapy utilizes QQMNCs with mainly angiogenic and anti-inflammatory phenotypes. QQMNCs promote the regenerative process in ischemic organs, and offer the advantages of relatively low cost and simple culture technique. EPC: endothelial progenitor cell, QQMNCs: quality and quantity culture system to obtain mononuclear cells.

Article Snippet: Moubarik et al. (2011) , 60-min transient occlusion of MCA in rats , Human UCB-derived EPCs (cultured EPCs from UCBMNCs; late EPCs, i.e., EOCs or ECFCs) , i.v. (24 hours after occlusion) from a femoral vein of Sprague-Dawley rat , Improved neurological deficits, increased capillary density, decreased apoptosis.

Techniques: Isolation

Journal: Neurologia medico-chirurgica

Article Title: Recent Progress in Endothelial Progenitor Cell Culture Systems: Potential for Stroke Therapy

doi: 10.2176/nmc.ra.2016-0027

Figure Lengend Snippet: Comparison of cell populations and characteristics of PBMNCs and QQMNCs. QQMNCs contain large EPCs, which are classified as definitive EPCs, with differentiating and vasculogenic capabilities. In addition, M2 macrophages are induced by anti-inflammatory cytokines and control adaptive immunity. Secreted anti-inflammatory cytokines promote and regulate type 2 immune responses, angiogenesis, and tissue repair. EPC: endothelial progenitor cell, PBMNC: peripheral blood mononuclear cell, QQMNCs: quality and quantity culture system to obtain mononuclear cells.

Article Snippet: Moubarik et al. (2011) , 60-min transient occlusion of MCA in rats , Human UCB-derived EPCs (cultured EPCs from UCBMNCs; late EPCs, i.e., EOCs or ECFCs) , i.v. (24 hours after occlusion) from a femoral vein of Sprague-Dawley rat , Improved neurological deficits, increased capillary density, decreased apoptosis.

Techniques: Comparison, Control

Journal: Neurologia medico-chirurgica

Article Title: Recent Progress in Endothelial Progenitor Cell Culture Systems: Potential for Stroke Therapy

doi: 10.2176/nmc.ra.2016-0027

Figure Lengend Snippet: Basic experiments of endothelial progenitor cell-based cell therapy for ischemic stroke

Article Snippet: Moubarik et al. (2011) , 60-min transient occlusion of MCA in rats , Human UCB-derived EPCs (cultured EPCs from UCBMNCs; late EPCs, i.e., EOCs or ECFCs) , i.v. (24 hours after occlusion) from a femoral vein of Sprague-Dawley rat , Improved neurological deficits, increased capillary density, decreased apoptosis.

Techniques: Cell Culture, Suspension

Journal: Bioengineered

Article Title: Endothelial progenitor cells-derived exosomes transfer microRNA-30e-5p to regulate Erastin-induced ferroptosis in human umbilical vein endothelial cells via the specificity protein 1/adenosine monophosphate-activated protein kinase axis

doi: 10.1080/21655979.2022.2025519

Figure Lengend Snippet: EPCs-Exos could be transferred to HUVECs. EPCs-Exos were extracted and identified. The uptake of EPCs-Exos by HUVECs was observed. (a) Morphological changes of EPCs were observed using an inverted microscope; (b) EPCs markers CD31 and CD34 were detected by immunofluorescence; (c) FITC-UEA-I and Dil-ac-LDL were detected by immunofluorescence; (d) The ultrastructure of Exos were observed using a TEM; (e) The diameter distribution and concentration of Exos were determined by NTA; (f) Exos mark proteins CD9, CD63, CD81, and Calnaxin were detected by WB; (g) The entry of PKH67-labeled Exos into HUVECs was detected by immunofluorescence. The cell experiments were repeated 3 times independently.

Article Snippet: Human umbilical vein blood-derived EPCs (HEPC, Cat No. CP-H164, Procell, Wuhan, Hubei, China) were washed twice with phosphate buffer saline (PBS) and centrifuged at 100 g. The cells were suspended in the endothelial basal growth medium supplemented with EGM-2 mV SingleQuots and 5% heat-inactivated fetal bovine serum (FBS) (Lonza, Basel, Switzerland).

Techniques: Inverted Microscopy, Immunofluorescence, Concentration Assay, Labeling

Journal: Bioengineered

Article Title: Endothelial progenitor cells-derived exosomes transfer microRNA-30e-5p to regulate Erastin-induced ferroptosis in human umbilical vein endothelial cells via the specificity protein 1/adenosine monophosphate-activated protein kinase axis

doi: 10.1080/21655979.2022.2025519

Figure Lengend Snippet: EPCs-Exos improved HUVECs ferroptosis and endothelial injury through regulating miR-30e-5p in HUVECs. HUVECs treated with Erastin were co-cultured with EPCs-Exos, EPCs-Exos inhibit-NC , or EPCs-Exos miR-inhibi , and the changes of HUVEC ferroptosis and injury were observed. (a) The expression of miR-30e-5p in HUVECs of each group was detected by RT-qPCR; (b) The expression of miR-30e-5p in EPCs was detected by RT-qPCR; (c) GSH content determination; (d) Lipid-ROS content determination; (e) MDA content determination; (f) Iron ion content determination; (g) Cell migration was detected by scratch assay; (h) Cell angiogenesis detection; (i) The expression of miR-30e-5p was detected by RT-qPCR after Exos were treated with RNase and detergent. The cell experiments were repeated 3 times independently. The data in the figure were all measurement data and were expressed as mean ± standard deviation. One-way ANOVA was used for data analysis. Tukey’s multiple comparisons test was used for the post hoc test. *** P < 0.001.

Article Snippet: Human umbilical vein blood-derived EPCs (HEPC, Cat No. CP-H164, Procell, Wuhan, Hubei, China) were washed twice with phosphate buffer saline (PBS) and centrifuged at 100 g. The cells were suspended in the endothelial basal growth medium supplemented with EGM-2 mV SingleQuots and 5% heat-inactivated fetal bovine serum (FBS) (Lonza, Basel, Switzerland).

Techniques: Cell Culture, Expressing, Quantitative RT-PCR, Migration, Wound Healing Assay, Standard Deviation

Journal: Bioengineered

Article Title: Endothelial progenitor cells-derived exosomes transfer microRNA-30e-5p to regulate Erastin-induced ferroptosis in human umbilical vein endothelial cells via the specificity protein 1/adenosine monophosphate-activated protein kinase axis

doi: 10.1080/21655979.2022.2025519

Figure Lengend Snippet: Overexpression of SP1 reversed the protective effect of EPCs-Exos on HUVECs against ferroptosis and endothelial injury. HUVECs were transfected with pcDNA3.1-SP1 and co-cultured with EPCs-Exos. The cell injury changes were observed. (a) The expression of SP1 was detected by RT-qPCR; (b) The protein level of SP1 was detected by WB; (c) GSH content determination; (d) Lipid-ROS content determination; (e) MDA content determination; (f) Iron ion content determination; (g) Cell migration was detected by scratch assay; (h) Cell angiogenesis detection; (i) The expression of miR-30e-5p was detected by RT-qPCR after Exos were treated with RNase and detergent. The cell experiments were repeated 3 times independently. The data in the figure were all measurement data and were expressed as mean ± standard deviation. One-way ANOVA was used for data analysis. Tukey’s multiple comparisons test was used for the post hoc test. *** P < 0.001. pc3.1-NC was pcDNA3.1-NC, pc3.1-SP1 was pcDNA3.1-SP1.

Article Snippet: Human umbilical vein blood-derived EPCs (HEPC, Cat No. CP-H164, Procell, Wuhan, Hubei, China) were washed twice with phosphate buffer saline (PBS) and centrifuged at 100 g. The cells were suspended in the endothelial basal growth medium supplemented with EGM-2 mV SingleQuots and 5% heat-inactivated fetal bovine serum (FBS) (Lonza, Basel, Switzerland).

Techniques: Over Expression, Transfection, Cell Culture, Expressing, Quantitative RT-PCR, Migration, Wound Healing Assay, Standard Deviation

Journal: Bioengineered

Article Title: Endothelial progenitor cells-derived exosomes transfer microRNA-30e-5p to regulate Erastin-induced ferroptosis in human umbilical vein endothelial cells via the specificity protein 1/adenosine monophosphate-activated protein kinase axis

doi: 10.1080/21655979.2022.2025519

Figure Lengend Snippet: EPCs-Exos promoted the AMPK pathway activation through miR-30e-5p targeting SP1. In HUVECs treated with Erastin and transfected with miR-30e-5p mimic/NC, and HUVECs co-cultured with EPCs-Exos, treated with Erastin, and transfected with pcDNA3.1-SP1, the protein phosphorylation of the AMPK pathway was observed. A: The expression of miR-30e-5p was detected by RT-qPCR; B: The protein levels of p-AMPK and AMPK were detected by WB. The cell experiments were repeated 3 times independently. The data in the figure were all measurement data and were expressed as mean ± standard deviation. One-way ANOVA was used for data analysis. Tukey’s multiple comparisons test was used for the post hoc test. *** P < 0.001. pc3.1-NC was pcDNA3.1-NC, pc3.1-SP1 was pcDNA3.1-SP1.

Article Snippet: Human umbilical vein blood-derived EPCs (HEPC, Cat No. CP-H164, Procell, Wuhan, Hubei, China) were washed twice with phosphate buffer saline (PBS) and centrifuged at 100 g. The cells were suspended in the endothelial basal growth medium supplemented with EGM-2 mV SingleQuots and 5% heat-inactivated fetal bovine serum (FBS) (Lonza, Basel, Switzerland).

Techniques: Activation Assay, Transfection, Cell Culture, Phospho-proteomics, Expressing, Quantitative RT-PCR, Standard Deviation

Journal: Bioengineered

Article Title: Endothelial progenitor cells-derived exosomes transfer microRNA-30e-5p to regulate Erastin-induced ferroptosis in human umbilical vein endothelial cells via the specificity protein 1/adenosine monophosphate-activated protein kinase axis

doi: 10.1080/21655979.2022.2025519

Figure Lengend Snippet: Mechanism diagram of EPCs-Exo uptake by HUVECs cells improving ferroptosis-induced by Erastin. HUVECs increased the level of intracellular GSH by inhibiting SP1 expression, reduced the levels of lipid ROS, MDA and iron ions, thus improving the decline of cell migration and angiogenesis caused by ferroptosis of HUVECs induced by Erastin.

Article Snippet: Human umbilical vein blood-derived EPCs (HEPC, Cat No. CP-H164, Procell, Wuhan, Hubei, China) were washed twice with phosphate buffer saline (PBS) and centrifuged at 100 g. The cells were suspended in the endothelial basal growth medium supplemented with EGM-2 mV SingleQuots and 5% heat-inactivated fetal bovine serum (FBS) (Lonza, Basel, Switzerland).

Techniques: Expressing, Migration