insulin growth factor 2 igf2 protein  (R&D Systems)

Journal: Stem Cell Research & Therapy

Article Title: Distinct muscle regenerative capacity of human induced pluripotent stem cell-derived mesenchymal stromal cells in Ullrich congenital muscular dystrophy model mice

doi: 10.1186/s13287-024-03951-6

Figure Lengend Snippet: Effects of IGF2 knockdown in XF-iMSCs on Col6a1 -KO/NSG MuSC differentiation. a mRNA expression of IGF2 and PXDN . The mRNA expression level of each gene was analyzed using RT-qPCR in Ad-MSCs, BM-MSCs, and XF-iMSCs. Levels are shown relative to those in XF-iMSCs. Data are shown as the mean ± SD. b Concentration of IGF2 in the culture supernatants of Ad-MSCs, BM-MSCs, and XF-iMSCs as obtained using ELISA. Data are shown as the mean ± SD. c mRNA expression level and concentration of IGF2 in the culture on co-culture day 3. Data are presented as the mean ± SD. mRNA expression levels are shown with levels relative to those in XF-iMSCs. d Representative immunofluorescence images of Col6a1 -KO/NSG mouse-derived MuSCs 3 days after single culture or co-culture with XF-iMSC, XF-iMSC_ IGF2 -KD, or XF-iMSC_mock. e Total number of DAPI + /hLamin A/C- mouse myogenic cells 3 days after co-culture. Data are expressed relative to XF-iMSCs and are presented as the mean ± SD of three independent experiments. f Percentage of Pax7 + /MyoD-, Pax7 + /MyoD + , and Pax7-/MyoD + cell populations 3 days after co-culture. Data from three independent experiments are shown as the mean ± SD. g mRNA expression level and concentration of IGF2 in the culture on co-culture day 6. Data are presented as the mean ± SD. mRNA expression levels are shown with levels relative to those in XF-iMSCs. h Representative immunofluorescence images of Col6a1 -KO/NSG mouse-derived MuSCs 6 days after single culture or co-culture with XF-iMSCs, XF-iMSC_ IGF2 -KD, or XF-iMSC_mock. i, j Area of MHC + myotubes ( i ) and number of MHC + myotubes ( j ) with four or more nuclei on day 6 after co-culture. Data are expressed relative to XF-iMSCs and are presented as the mean ± SD of three independent experiments. co-cluture experiment: n = 6 (XF-iMSCs, XF-iMSC_ IGF2-KD and XF-iMSC_mock), n = 3 (single culture). qPCR and ELISA experiment: n = 2 (Ad-MSCs, BM-MSCs, XF-iMSCs)

Article Snippet: Insulin growth factor 2 (IGF2) protein was quantified using a Human IGF-II/IGF2 Quantizing ELISA kit (DG200, R&D Systems, Minneapolis, MSP, USA).

Techniques: Knockdown, Expressing, Quantitative RT-PCR, Concentration Assay, Enzyme-linked Immunosorbent Assay, Co-Culture Assay, Immunofluorescence, Derivative Assay

Journal: Stem Cell Research & Therapy

Article Title: Distinct muscle regenerative capacity of human induced pluripotent stem cell-derived mesenchymal stromal cells in Ullrich congenital muscular dystrophy model mice

doi: 10.1186/s13287-024-03951-6

Figure Lengend Snippet: Effects of IGF2 supplementation on Col6a1 -KO/NSG MuSC differentiation. a Representative immunofluorescence images of Col6a1 -KO/NSG mouse-derived MuSCs 3 days after IGF2 supplementation (IGF2-treated) or without IGF2 treatment (IGF2-untreated). b Total number of DAPI + /hLamin A/C- mouse myogenic cells after 3 days of culture. Data are expressed relative to IGF2-untreated and are presented as the mean ± SD of three independent experiments. c Percentage of Pax7 + /MyoD-, Pax7 + /MyoD + , and Pax7-/MyoD + cell populations after 3 days of culture. Data from three independent experiments are shown as the mean ± SD. d Representative immunofluorescence images of Col6a1 -KO/NSG mouse-derived MuSCs 6 days after IGF2 supplementation (IGF2-treated) or without IGF2 treatment (IGF2-untreated). e, f Area of MHC + myotubes ( e ) and number of MHC + myotubes with two or more nuclei ( f ) 6 days after co-culture. Data are expressed relative to IGF2-untreated and are presented as the mean ± SD of three independent experiments. * p < 0.05. n = 3 (IGF2 treated, IGF2 untreated)

Article Snippet: Insulin growth factor 2 (IGF2) protein was quantified using a Human IGF-II/IGF2 Quantizing ELISA kit (DG200, R&D Systems, Minneapolis, MSP, USA).

Techniques: Immunofluorescence, Derivative Assay, Co-Culture Assay

Journal: Stem Cells Translational Medicine

Article Title: Enforced Expression of HOXB4 in Human Embryonic Stem Cells Enhances the Production of Hematopoietic Progenitors but Has No Effect on the Maturation of Red Blood Cells

doi: 10.5966/sctm.2015-0324

Figure Lengend Snippet: Activation of HOXB4 enhanced the production of hematopoietic progenitors. Tamoxifen (labeled + or −) was added to cultures to activate HOXB4 in H1 human embryonic stem cells (hESCs) expressing HOXB4-ER T2 (labeled HOXB4) or parental H1 hESCs (labeled control) between days 0 and 10 (A) and then assessed for CFU-C activity (B) and expression of hematopoietic progenitor markers, CD34 and CD43, by flow cytometry (C, D) . Data were generated from four independent experiments, with error bars representing SEM. ∗, p = .03; ∗∗, p = .04. Abbreviations: 4OHT, tamoxifen; BFU-E, burst-forming unit erythroid; BMP4, bone morphogenetic protein 4; C, cell; CFU, colony-forming unit; FGF a, fibroblast growth factor-α; GM, granulocyte macrophage; GSK, glycogen synthase kinase; IBMX, isobutylmethylxanthine; IGF2, insulin-like growth factor 2; M, macrophage; Q, quartile; SCF, stem cell factor; TPO, thrombopoietin; VEGF, vascular endothelial growth factor.

Article Snippet: After centrifugation, the cells were replated at 2 × 10 5 cells per well (six-well plates; Corning, Corning, NY, http://www.corning.com ) in 3 ml of Stemline II, with the addition of BMP4 20 ng/ml, VEGF 30 ng/ml, FGF-α 10 ng/ml, SCF 30 ng/ml, insulin-like growth factor 2 (IGF2) 10 ng/ml (catalog no. 292-G2; R&D Systems), thrombopoietin (TPO) 10 ng/ml (catalog no. 288-TPN-25; R&D Systems), heparin 5 μg/ml (catalog no. H3149; Sigma-Aldrich), and isobutylmethylxanthine (IBMX) 100 μM (catalog no. I5879; Sigma-Aldrich).

Techniques: Activation Assay, Labeling, Expressing, Control, Activity Assay, Flow Cytometry, Generated

Journal: Stem Cells Translational Medicine

Article Title: Enforced Expression of HOXB4 in Human Embryonic Stem Cells Enhances the Production of Hematopoietic Progenitors but Has No Effect on the Maturation of Red Blood Cells

doi: 10.5966/sctm.2015-0324

Figure Lengend Snippet: Activation of HOXB4 resulted in a modest increase in the proportion of immature CD235a + /CD71 + erythroid cells. HOXB4 was activated with 4OHT from days 10 to 24 (A) . Production of erythroid cells was monitored by expression of CD235a and CD71 by flow cytometry at day 24 (B–D) . Data were generated from four independent experiments with error bars representing the SEM. ∗, p = .05. Abbreviations: 4OHT, tamoxifen; BMP4, bone morphogenetic protein 4; C, cell; CFU, colony-forming unit; EPO, erythropoietin; FGF a, fibroblast growth factor-α; FLT3L, FMS-like tyrosine kinase receptor 3 ligand; GSK, glycogen synthase kinase; IBMX, isobutylmethylxanthine; IGF1, IGF2, insulin-like growth factor 1, 2; IL, interleukin; Q, quartile; SCF, stem cell factor; TPO, thrombopoietin; VEGF, vascular endothelial growth factor.

Article Snippet: After centrifugation, the cells were replated at 2 × 10 5 cells per well (six-well plates; Corning, Corning, NY, http://www.corning.com ) in 3 ml of Stemline II, with the addition of BMP4 20 ng/ml, VEGF 30 ng/ml, FGF-α 10 ng/ml, SCF 30 ng/ml, insulin-like growth factor 2 (IGF2) 10 ng/ml (catalog no. 292-G2; R&D Systems), thrombopoietin (TPO) 10 ng/ml (catalog no. 288-TPN-25; R&D Systems), heparin 5 μg/ml (catalog no. H3149; Sigma-Aldrich), and isobutylmethylxanthine (IBMX) 100 μM (catalog no. I5879; Sigma-Aldrich).

Techniques: Activation Assay, Expressing, Flow Cytometry, Generated