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Vasculogenic conditioning of peripheral blood mononuclear cells promotes endothelial progenitor cell expansion and phenotype transition of anti-inflammatory macrophage and T lymphocyte to cells with regenerative potential.

Masuda H, Tanaka R, Fujimura S, Ishikawa M, Akimaru H, Shizuno T, Sato A, Okada Y, Iida Y, Itoh J, Itoh Y, Kamiguchi H, Kawamoto A, Asahara T - J Am Heart Assoc (2014)

Bottom Line: The resulting cells (QQMNCs) in EPC colony-forming assay generated significantly more definitive EPC colonies than PBMNCs.Using murine ischemic hindlimb models, the efficacy of QQMNC intramuscular transplantation (Tx) was compared to that of PBMNCTx, cultured "early EPC" Tx (eEPCTx), and granulocyte colony-stimulating factor mobilized CD34(+) cell Tx (GmCD34Tx).Laser Doppler imaging revealed the blood perfusion recovery in ischemic hindlimbs after QQMNCTx superior to after PBMNCTx and eEPCTx, but also earlier than after GmCD34Tx.

View Article: PubMed Central - PubMed

Affiliation: Department of Regenerative Medicine Science, Tokai University School of Medicine, Isehara, Japan (H.M., T.S., A.S., T.A.).

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In vitro angiogenic assay of HUVECs cocultured with QQMNCs. A, Features of tubes formed by HUVECs. B, The bar graphs represent the number of tubes counted under ×2 HPF. *P<0.05 versus HUVEC; ##P<0.01 versus HUVEC+PBMNC. C, Incorporation of acLDL‐DiI uptaking PBMNCs or QQMNCs into tubes of HUVECs. Arrow heads indicate PBMNCs or QQMNCs that are labeled with acLDL‐DiI and incorporated into tubes formed by HUVECs. (a) HUVEC alone, (b) HUVEC+PBMNC, and (c) HUVEC+QQMNC in (A and C). D, The bar graphs show numbers of incorporated PBMNCs or QQMNCs, counted under ×4 HPF. ***P<0.001. Each graph column represents a mean±SE. N=10 wells/group. acLDL‐DiI indicates acetylated low density lipoprotein, labeled with 1,1′‐dioctadecyl‐3,3,3′,3′‐tetramethyl‐indocarbocyanine perchlorate; HPF, high power field; HUVECs, human umbilical vein endothelial cells; PBMNCs, peripheral blood mononuclear cells; QQMNCs, quality and quantity control culture of mononuclear cells.
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fig04: In vitro angiogenic assay of HUVECs cocultured with QQMNCs. A, Features of tubes formed by HUVECs. B, The bar graphs represent the number of tubes counted under ×2 HPF. *P<0.05 versus HUVEC; ##P<0.01 versus HUVEC+PBMNC. C, Incorporation of acLDL‐DiI uptaking PBMNCs or QQMNCs into tubes of HUVECs. Arrow heads indicate PBMNCs or QQMNCs that are labeled with acLDL‐DiI and incorporated into tubes formed by HUVECs. (a) HUVEC alone, (b) HUVEC+PBMNC, and (c) HUVEC+QQMNC in (A and C). D, The bar graphs show numbers of incorporated PBMNCs or QQMNCs, counted under ×4 HPF. ***P<0.001. Each graph column represents a mean±SE. N=10 wells/group. acLDL‐DiI indicates acetylated low density lipoprotein, labeled with 1,1′‐dioctadecyl‐3,3,3′,3′‐tetramethyl‐indocarbocyanine perchlorate; HPF, high power field; HUVECs, human umbilical vein endothelial cells; PBMNCs, peripheral blood mononuclear cells; QQMNCs, quality and quantity control culture of mononuclear cells.

Mentions: Using an in vitro Matrigel assay, we found that QQMNCs promoted tube formation of cocultured HUVECs for 12 hours, but PBMNCs did not (tube counts/×2 HPF=63.3±1.43 for HUVEC+QQMNC versus 55.1±1.45 for HUVEC+PBMNC or 55.3±1.39 for HUVEC alone; Figure 4A and 4B).


Vasculogenic conditioning of peripheral blood mononuclear cells promotes endothelial progenitor cell expansion and phenotype transition of anti-inflammatory macrophage and T lymphocyte to cells with regenerative potential.

Masuda H, Tanaka R, Fujimura S, Ishikawa M, Akimaru H, Shizuno T, Sato A, Okada Y, Iida Y, Itoh J, Itoh Y, Kamiguchi H, Kawamoto A, Asahara T - J Am Heart Assoc (2014)

In vitro angiogenic assay of HUVECs cocultured with QQMNCs. A, Features of tubes formed by HUVECs. B, The bar graphs represent the number of tubes counted under ×2 HPF. *P<0.05 versus HUVEC; ##P<0.01 versus HUVEC+PBMNC. C, Incorporation of acLDL‐DiI uptaking PBMNCs or QQMNCs into tubes of HUVECs. Arrow heads indicate PBMNCs or QQMNCs that are labeled with acLDL‐DiI and incorporated into tubes formed by HUVECs. (a) HUVEC alone, (b) HUVEC+PBMNC, and (c) HUVEC+QQMNC in (A and C). D, The bar graphs show numbers of incorporated PBMNCs or QQMNCs, counted under ×4 HPF. ***P<0.001. Each graph column represents a mean±SE. N=10 wells/group. acLDL‐DiI indicates acetylated low density lipoprotein, labeled with 1,1′‐dioctadecyl‐3,3,3′,3′‐tetramethyl‐indocarbocyanine perchlorate; HPF, high power field; HUVECs, human umbilical vein endothelial cells; PBMNCs, peripheral blood mononuclear cells; QQMNCs, quality and quantity control culture of mononuclear cells.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4309104&req=5

fig04: In vitro angiogenic assay of HUVECs cocultured with QQMNCs. A, Features of tubes formed by HUVECs. B, The bar graphs represent the number of tubes counted under ×2 HPF. *P<0.05 versus HUVEC; ##P<0.01 versus HUVEC+PBMNC. C, Incorporation of acLDL‐DiI uptaking PBMNCs or QQMNCs into tubes of HUVECs. Arrow heads indicate PBMNCs or QQMNCs that are labeled with acLDL‐DiI and incorporated into tubes formed by HUVECs. (a) HUVEC alone, (b) HUVEC+PBMNC, and (c) HUVEC+QQMNC in (A and C). D, The bar graphs show numbers of incorporated PBMNCs or QQMNCs, counted under ×4 HPF. ***P<0.001. Each graph column represents a mean±SE. N=10 wells/group. acLDL‐DiI indicates acetylated low density lipoprotein, labeled with 1,1′‐dioctadecyl‐3,3,3′,3′‐tetramethyl‐indocarbocyanine perchlorate; HPF, high power field; HUVECs, human umbilical vein endothelial cells; PBMNCs, peripheral blood mononuclear cells; QQMNCs, quality and quantity control culture of mononuclear cells.
Mentions: Using an in vitro Matrigel assay, we found that QQMNCs promoted tube formation of cocultured HUVECs for 12 hours, but PBMNCs did not (tube counts/×2 HPF=63.3±1.43 for HUVEC+QQMNC versus 55.1±1.45 for HUVEC+PBMNC or 55.3±1.39 for HUVEC alone; Figure 4A and 4B).

Bottom Line: The resulting cells (QQMNCs) in EPC colony-forming assay generated significantly more definitive EPC colonies than PBMNCs.Using murine ischemic hindlimb models, the efficacy of QQMNC intramuscular transplantation (Tx) was compared to that of PBMNCTx, cultured "early EPC" Tx (eEPCTx), and granulocyte colony-stimulating factor mobilized CD34(+) cell Tx (GmCD34Tx).Laser Doppler imaging revealed the blood perfusion recovery in ischemic hindlimbs after QQMNCTx superior to after PBMNCTx and eEPCTx, but also earlier than after GmCD34Tx.

View Article: PubMed Central - PubMed

Affiliation: Department of Regenerative Medicine Science, Tokai University School of Medicine, Isehara, Japan (H.M., T.S., A.S., T.A.).

Show MeSH
Related in: MedlinePlus