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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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Related in: MedlinePlus

Characteristics of QQMNCs versus PBMNCs. A, The graph shows total cell counts of PBMNCs isolated from 100 mL of PB and the respective QQMNC counts. B, The left graph indicates linear regression analysis between the cell‐count ratio of QQMNCs versus PBMNCs (2×106 cells/well) and PBMNC counts isolated from 100 mL of PB. The right indicates linear regression analysis of cell counts between PBMNCs and QQMNCs per 100 mL of PB. C, Representative pictures of pEPC‐CFU and dEPC‐CFU. Scale bar=500 μm. D, The left and middle graphs are EPC‐CFU counts generated from PBMNCs or QQMNCs per dish (2×105 cells/dish) and in 100 mL of PB. The right graph shows the percentage of each EPC‐CFU count versus total EPC‐CFU count per dish. Each column in the graph represents a mean±SE. The white and gray areas in the bar graphs indicate the values of pEPC‐CFU and dEPC‐CFU. E, Linear regression graphs on the interrelation of each EPC‐CFU count per dish in QQMNCs to that in PBMNCs. *P<0.05; ***P<0.001. N=18 volunteers. F, Comparison of EPC colony forming activities of post‐QQ cultured cells among CD34+ cells, CD34‐MNCs, and their repopulated cells. QQ‐34‐MNC: QQ cultured cells of CD34‐MNCs (2×106 cells/2 mL of QQ culture medium), QQ‐34+: QQ cultured cells of CD34+ cells alone (4×103 cells/2 mL of QQ culture medium); QQ‐34+/34‐MNC: QQ cultured cells of CD34+ cells repopulated CD34‐MNCs (4×103 cells for CD34+ cells with 2×106 cells for CD34‐MNCs/2 mL of QQ culture medium). *P<0.05; **P<0.01; ***P<0.001 versus QQ‐34‐MNC. #P<0.05; ###P<0.001 versus QQ‐34+. Each column in the graph represents a mean±SE. N=3 volunteers. Aliquots of each cell cultured in the equal volume of QQ culture medium were applied to EPC‐CFA; the aliquots were respectively seeded at 2×105 cells/dish (3 dishes each for 3 volunteers) for QQ‐34‐MNC and QQ‐34+/34‐MNC, and at the ratio of 2×105 cells to the QQ‐34+/34‐MNCs for QQ‐34+. dEPC‐CFU indicates definitive endothelial progenitor cells colony‐forming units; PBMNCs, peripheral blood mononuclear cells; pEPC‐CFU, primitive endothelial progenitor cells colony‐forming units; QQMNCs, quality and quantity control culture of mononuclear cells.
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fig01: Characteristics of QQMNCs versus PBMNCs. A, The graph shows total cell counts of PBMNCs isolated from 100 mL of PB and the respective QQMNC counts. B, The left graph indicates linear regression analysis between the cell‐count ratio of QQMNCs versus PBMNCs (2×106 cells/well) and PBMNC counts isolated from 100 mL of PB. The right indicates linear regression analysis of cell counts between PBMNCs and QQMNCs per 100 mL of PB. C, Representative pictures of pEPC‐CFU and dEPC‐CFU. Scale bar=500 μm. D, The left and middle graphs are EPC‐CFU counts generated from PBMNCs or QQMNCs per dish (2×105 cells/dish) and in 100 mL of PB. The right graph shows the percentage of each EPC‐CFU count versus total EPC‐CFU count per dish. Each column in the graph represents a mean±SE. The white and gray areas in the bar graphs indicate the values of pEPC‐CFU and dEPC‐CFU. E, Linear regression graphs on the interrelation of each EPC‐CFU count per dish in QQMNCs to that in PBMNCs. *P<0.05; ***P<0.001. N=18 volunteers. F, Comparison of EPC colony forming activities of post‐QQ cultured cells among CD34+ cells, CD34‐MNCs, and their repopulated cells. QQ‐34‐MNC: QQ cultured cells of CD34‐MNCs (2×106 cells/2 mL of QQ culture medium), QQ‐34+: QQ cultured cells of CD34+ cells alone (4×103 cells/2 mL of QQ culture medium); QQ‐34+/34‐MNC: QQ cultured cells of CD34+ cells repopulated CD34‐MNCs (4×103 cells for CD34+ cells with 2×106 cells for CD34‐MNCs/2 mL of QQ culture medium). *P<0.05; **P<0.01; ***P<0.001 versus QQ‐34‐MNC. #P<0.05; ###P<0.001 versus QQ‐34+. Each column in the graph represents a mean±SE. N=3 volunteers. Aliquots of each cell cultured in the equal volume of QQ culture medium were applied to EPC‐CFA; the aliquots were respectively seeded at 2×105 cells/dish (3 dishes each for 3 volunteers) for QQ‐34‐MNC and QQ‐34+/34‐MNC, and at the ratio of 2×105 cells to the QQ‐34+/34‐MNCs for QQ‐34+. dEPC‐CFU indicates definitive endothelial progenitor cells colony‐forming units; PBMNCs, peripheral blood mononuclear cells; pEPC‐CFU, primitive endothelial progenitor cells colony‐forming units; QQMNCs, quality and quantity control culture of mononuclear cells.

Mentions: The fold increase of QQMNCs to PBMNCs per well declined in the whole subjects with an average of 0.54‐fold (Table 11). The calculated total QQMNCs derived from 100 mL of PB decreased from original cells (cell counts×105=831.3±75.3 to 399.2±43.1), on average, by 0.48‐fold (Figure 1A; Table 11).


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)

Characteristics of QQMNCs versus PBMNCs. A, The graph shows total cell counts of PBMNCs isolated from 100 mL of PB and the respective QQMNC counts. B, The left graph indicates linear regression analysis between the cell‐count ratio of QQMNCs versus PBMNCs (2×106 cells/well) and PBMNC counts isolated from 100 mL of PB. The right indicates linear regression analysis of cell counts between PBMNCs and QQMNCs per 100 mL of PB. C, Representative pictures of pEPC‐CFU and dEPC‐CFU. Scale bar=500 μm. D, The left and middle graphs are EPC‐CFU counts generated from PBMNCs or QQMNCs per dish (2×105 cells/dish) and in 100 mL of PB. The right graph shows the percentage of each EPC‐CFU count versus total EPC‐CFU count per dish. Each column in the graph represents a mean±SE. The white and gray areas in the bar graphs indicate the values of pEPC‐CFU and dEPC‐CFU. E, Linear regression graphs on the interrelation of each EPC‐CFU count per dish in QQMNCs to that in PBMNCs. *P<0.05; ***P<0.001. N=18 volunteers. F, Comparison of EPC colony forming activities of post‐QQ cultured cells among CD34+ cells, CD34‐MNCs, and their repopulated cells. QQ‐34‐MNC: QQ cultured cells of CD34‐MNCs (2×106 cells/2 mL of QQ culture medium), QQ‐34+: QQ cultured cells of CD34+ cells alone (4×103 cells/2 mL of QQ culture medium); QQ‐34+/34‐MNC: QQ cultured cells of CD34+ cells repopulated CD34‐MNCs (4×103 cells for CD34+ cells with 2×106 cells for CD34‐MNCs/2 mL of QQ culture medium). *P<0.05; **P<0.01; ***P<0.001 versus QQ‐34‐MNC. #P<0.05; ###P<0.001 versus QQ‐34+. Each column in the graph represents a mean±SE. N=3 volunteers. Aliquots of each cell cultured in the equal volume of QQ culture medium were applied to EPC‐CFA; the aliquots were respectively seeded at 2×105 cells/dish (3 dishes each for 3 volunteers) for QQ‐34‐MNC and QQ‐34+/34‐MNC, and at the ratio of 2×105 cells to the QQ‐34+/34‐MNCs for QQ‐34+. dEPC‐CFU indicates definitive endothelial progenitor cells colony‐forming units; PBMNCs, peripheral blood mononuclear cells; pEPC‐CFU, primitive endothelial progenitor cells colony‐forming units; QQMNCs, quality and quantity control culture of mononuclear cells.
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Related In: Results  -  Collection

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fig01: Characteristics of QQMNCs versus PBMNCs. A, The graph shows total cell counts of PBMNCs isolated from 100 mL of PB and the respective QQMNC counts. B, The left graph indicates linear regression analysis between the cell‐count ratio of QQMNCs versus PBMNCs (2×106 cells/well) and PBMNC counts isolated from 100 mL of PB. The right indicates linear regression analysis of cell counts between PBMNCs and QQMNCs per 100 mL of PB. C, Representative pictures of pEPC‐CFU and dEPC‐CFU. Scale bar=500 μm. D, The left and middle graphs are EPC‐CFU counts generated from PBMNCs or QQMNCs per dish (2×105 cells/dish) and in 100 mL of PB. The right graph shows the percentage of each EPC‐CFU count versus total EPC‐CFU count per dish. Each column in the graph represents a mean±SE. The white and gray areas in the bar graphs indicate the values of pEPC‐CFU and dEPC‐CFU. E, Linear regression graphs on the interrelation of each EPC‐CFU count per dish in QQMNCs to that in PBMNCs. *P<0.05; ***P<0.001. N=18 volunteers. F, Comparison of EPC colony forming activities of post‐QQ cultured cells among CD34+ cells, CD34‐MNCs, and their repopulated cells. QQ‐34‐MNC: QQ cultured cells of CD34‐MNCs (2×106 cells/2 mL of QQ culture medium), QQ‐34+: QQ cultured cells of CD34+ cells alone (4×103 cells/2 mL of QQ culture medium); QQ‐34+/34‐MNC: QQ cultured cells of CD34+ cells repopulated CD34‐MNCs (4×103 cells for CD34+ cells with 2×106 cells for CD34‐MNCs/2 mL of QQ culture medium). *P<0.05; **P<0.01; ***P<0.001 versus QQ‐34‐MNC. #P<0.05; ###P<0.001 versus QQ‐34+. Each column in the graph represents a mean±SE. N=3 volunteers. Aliquots of each cell cultured in the equal volume of QQ culture medium were applied to EPC‐CFA; the aliquots were respectively seeded at 2×105 cells/dish (3 dishes each for 3 volunteers) for QQ‐34‐MNC and QQ‐34+/34‐MNC, and at the ratio of 2×105 cells to the QQ‐34+/34‐MNCs for QQ‐34+. dEPC‐CFU indicates definitive endothelial progenitor cells colony‐forming units; PBMNCs, peripheral blood mononuclear cells; pEPC‐CFU, primitive endothelial progenitor cells colony‐forming units; QQMNCs, quality and quantity control culture of mononuclear cells.
Mentions: The fold increase of QQMNCs to PBMNCs per well declined in the whole subjects with an average of 0.54‐fold (Table 11). The calculated total QQMNCs derived from 100 mL of PB decreased from original cells (cell counts×105=831.3±75.3 to 399.2±43.1), on average, by 0.48‐fold (Figure 1A; Table 11).

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