Journal: Stem Cells International

Article Title: Transplantation of Rat Mesenchymal Stem Cells Overexpressing Hypoxia-Inducible Factor 2 α Improves Blood Perfusion and Arteriogenesis in a Rat Hindlimb Ischemia Model

doi: 10.1155/2017/3794817

Figure Lengend Snippet: HIF-2 α was overexpressed in rMSCs and rMSCs HIF-2 α showed stem cell characteristics. (a) Representative fluorescence images showing rMSCs transduced with 0, 25, 50, or 100 MOI recombinant lentivirus expressing HIF-2 α . The images were collected 48 hours after transduction. (b) The dose of MOI 100 resulted in >80% transduction efficiency. (c) Representative image of Western blot showing HIF-2 α expression in rMSCs transduced with different doses of recombinant lentivirus. (d) Representative image of flow cytometry analysis showing abundant expression of CD29 and absence of CD45 on rMSCs HIF-2 α . (e) Alizarin red S staining shows calcium deposit in osteoblasts differentiated from rMSCs HIF-2 α . (f) Adipocytes differentiated from rMSCs HIF-2 α shows positive oil red O staining for lipid droplets. (g) Toluidine blue staining shows positive glycosaminoglycan matrix production from chondrocytes differentiated from rMSCs HIF-2 α .

Article Snippet: Rat mesenchymal stem cells (rMSCs) were purchased from Cyagen Biosciences Inc., China, and cultured in F344 Rat Mesenchymal Stem Cell Basal media supplemented with 10% Mesenchymal Stem Cell-Qualified Fetal Bovine Serum (Cyagen Biosciences Inc., China), 1% penicillin-streptomycin, and 1% glutamine (Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Fluorescence, Transduction, Recombinant, Expressing, Western Blot, Flow Cytometry, Staining

Journal: Stem Cells International

Article Title: Transplantation of Rat Mesenchymal Stem Cells Overexpressing Hypoxia-Inducible Factor 2 α Improves Blood Perfusion and Arteriogenesis in a Rat Hindlimb Ischemia Model

doi: 10.1155/2017/3794817

Figure Lengend Snippet: Transplantation of rMSCs HIF-2 α improved blood perfusion and increased neoangiogenesis. (a) Representative LDP images of the ischemic and nonischemic limbs. The images were collected 14 days after the transplantation. (b) Representative photos of the ischemic and nonischemic limbs. The photos were obtained 14 days after the transplantation. The sham group photo shows normal rat feet; the PBS group photo shows a loss of the big toe in the ischemic foot; the rMSC group photo shows gangrene in the ischemic foot; the rMSCs HIF-2 α group photo shows relatively normal rat feet. (c) The rMSCs HIF-2 α group showed significantly higher LDPI index than the rMSC and PBS groups. LDPI index was calculated as the ratio of the perfusion in the ischemic limb to the perfusion in the nonischemic limb. ∗ indicates significant difference of the rMSCs HIF-2 α group versus rMSC and PBS groups, P < 0.05. # indicates significant difference of the rMSCs HIF-2 α group versus rMSC and PBS groups, P < 0.01. (d) The rMSCs HIF-2 α group showed significantly lower mobility damage score of the ischemic limb than the PBS groups. ∗ indicates significant difference of the rMSCs HIF-2 α group versus the PBS group, P < 0.05. (e) Representative images of staining for alkaline phosphatase and α -smooth muscle actin (SMA). (f) Capillary density in the rMSCs and rMSCs HIF-2 α groups was significantly higher than that in the PBS group. ∗ indicates significant difference of the indicated groups versus PBS group, P < 0.05. (g) Arteriole density in the rMSCs and rMSCs HIF-2 α groups was significantly higher than that in the PBS group. ∗ indicates significant difference of the indicated groups versus PBS group, P < 0.05. # indicates significant difference of the indicated group versus PBS group, P < 0.01.

Article Snippet: Rat mesenchymal stem cells (rMSCs) were purchased from Cyagen Biosciences Inc., China, and cultured in F344 Rat Mesenchymal Stem Cell Basal media supplemented with 10% Mesenchymal Stem Cell-Qualified Fetal Bovine Serum (Cyagen Biosciences Inc., China), 1% penicillin-streptomycin, and 1% glutamine (Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Transplantation Assay, Staining

Journal: Stem Cells International

Article Title: Transplantation of Rat Mesenchymal Stem Cells Overexpressing Hypoxia-Inducible Factor 2 α Improves Blood Perfusion and Arteriogenesis in a Rat Hindlimb Ischemia Model

doi: 10.1155/2017/3794817

Figure Lengend Snippet: Transplantation of rMSCs HIF-2 α increased the number of vessel branches. (a) Representative images of synchrotron radiation microangiography. (b) Angiographic score in the rMSC and rMSC HIF-2 α groups was significantly higher than that in the PBS group. ∗ indicates significant difference of the indicated groups versus PBS group, P < 0.05. (c) The number of vessel branches in the rMSC HIF-2 α group was significantly higher than that in the rMSCs and PBS groups. ∗ indicates significant difference of the indicated group versus rMSCs and PBS groups, P < 0.05.

Article Snippet: Rat mesenchymal stem cells (rMSCs) were purchased from Cyagen Biosciences Inc., China, and cultured in F344 Rat Mesenchymal Stem Cell Basal media supplemented with 10% Mesenchymal Stem Cell-Qualified Fetal Bovine Serum (Cyagen Biosciences Inc., China), 1% penicillin-streptomycin, and 1% glutamine (Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Transplantation Assay

Journal: Stem Cells International

Article Title: Transplantation of Rat Mesenchymal Stem Cells Overexpressing Hypoxia-Inducible Factor 2 α Improves Blood Perfusion and Arteriogenesis in a Rat Hindlimb Ischemia Model

doi: 10.1155/2017/3794817

Figure Lengend Snippet: Cell transplantation increased the expression of growth factors and HIF-2 α targeting molecules in the muscle tissue of the ischemic limb and cause dynamic change in serum angiopoietin. (a) Representative image of RT-PCR showing that the mRNA levels of VEGF, bFGF, and SDF1 increased and Bax mRNA level reduced 7 days after transplantation of rMSCs or rMSCs HIF-2 α . (b and c) Representative images of Western blot showing that the expression of DII4, NICD, Hey1, Hes1, VEGFR2, VEGF-A, NRP-1, and Tie-2 increased 14 days after transplantation of rMSCs or rMSCs HIF-2 α . (d) ELISA analysis of serum Ang1. (e) ELISA analysis of serum Ang2.

Article Snippet: Rat mesenchymal stem cells (rMSCs) were purchased from Cyagen Biosciences Inc., China, and cultured in F344 Rat Mesenchymal Stem Cell Basal media supplemented with 10% Mesenchymal Stem Cell-Qualified Fetal Bovine Serum (Cyagen Biosciences Inc., China), 1% penicillin-streptomycin, and 1% glutamine (Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Transplantation Assay, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Stem Cells International

Article Title: Transplantation of Rat Mesenchymal Stem Cells Overexpressing Hypoxia-Inducible Factor 2 α Improves Blood Perfusion and Arteriogenesis in a Rat Hindlimb Ischemia Model

doi: 10.1155/2017/3794817

Figure Lengend Snippet: Effect of HIF-2 α overexpression on rMSC behavior. (a) HIF-2 α overexpression did not affect rMSC proliferation. (b) HIF-2 α overexpression increased rMSC migration under hypoxia (5% O 2 ) condition. The representative images of scratched cell layer were collected after 24 hours of incubation. ∗ indicates significant difference of the indicated group versus rMSCs group under the same condition, P < 0.05. (c) HIF-2 α overexpression increased rMSC tube formation in matrigel under hypoxia (5% O 2 ) condition. The representative images of tube formation in matrigel were collected after 24 hours of incubation. ∗ indicates significant difference of the indicated group versus rMSCs group under the same condition, P < 0.05. (d) Coculture with rMSCs HIF-2 α significantly increased the invasion of endothelial cells under hypoxia condition. The representative images of invaded endothelial cells were collected after 24 hours of incubation. ∗ indicates significant difference of the indicated group versus rMSCs group under the same condition, P < 0.05.

Article Snippet: Rat mesenchymal stem cells (rMSCs) were purchased from Cyagen Biosciences Inc., China, and cultured in F344 Rat Mesenchymal Stem Cell Basal media supplemented with 10% Mesenchymal Stem Cell-Qualified Fetal Bovine Serum (Cyagen Biosciences Inc., China), 1% penicillin-streptomycin, and 1% glutamine (Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Over Expression, Migration, Incubation