TY - JOUR
T1 - Characteristics and Function of Cryopreserved Bone Marrow-Derived Endothelial Progenitor Cells
AU - Mieno, Shigetoshi
AU - Clements, Richard T.
AU - Boodhwani, Munir
AU - Sodha, Neel R.
AU - Ramlawi, Basel
AU - Bianchi, Cesario
AU - Sellke, Frank W.
N1 - Copyright:
Copyright 2009 Elsevier B.V., All rights reserved.
PY - 2008/4
Y1 - 2008/4
N2 - Background: This study examined ex vivo expansion of bone marrow-derived endothelial progenitor cell (EPC) from cryopreserved bone marrow-derived mononuclear cells, and evaluated proliferation and migration function of the cryopreserved EPC (Cryo-EPC). Methods: Bone marrow samples were taken from swine iliac bone (n = 6). Isolated bone marrow-derived mononuclear cells were cultured or cryopreserved at -80°C for 2 to 3 months. After cell culture for 4 days, attached cells, EPCs with or without cryopreservation, were collected. Direct fluorescent staining by acetylated low-density lipoprotein, isolectin B4, and 4′,6-diamidino-2-phenylindole were performed to confirm the attached cells as EPC. Endothelial progenitor cell proliferation by vascular endothelial growth factor was evaluated by the tetrazolium method. Endothelial progenitor cell migration in response to stromal-derived factor-1α was also evaluated by using a Boyden chamber assay. Results: The percentage of cells positively stained by direct fluorescent staining by acetylated low-density lipoprotein and isolectin B4 was similar between fresh and Cryo-EPC (EPC = 96.0 ± 0.42 versus Cryo-EPC = 95.2 ± 1.2; p = 0.21). Vascular endothelial growth factor increased proliferation activity in fresh and Cryo-EPC (p < 0.01). Stromal-derived factor-1α increased migration activity in fresh and Cryo-EPC (p < 0.01). There was no difference in proliferation and migration activity between fresh and Cryo-EPC. Conclusions: Ex vivo expansion by cell culture was a useful method for collection of bone marrow-derived EPC from cryopreserved mononuclear cells. Proliferation and migration function of EPC is preserved after cryopreservation.
AB - Background: This study examined ex vivo expansion of bone marrow-derived endothelial progenitor cell (EPC) from cryopreserved bone marrow-derived mononuclear cells, and evaluated proliferation and migration function of the cryopreserved EPC (Cryo-EPC). Methods: Bone marrow samples were taken from swine iliac bone (n = 6). Isolated bone marrow-derived mononuclear cells were cultured or cryopreserved at -80°C for 2 to 3 months. After cell culture for 4 days, attached cells, EPCs with or without cryopreservation, were collected. Direct fluorescent staining by acetylated low-density lipoprotein, isolectin B4, and 4′,6-diamidino-2-phenylindole were performed to confirm the attached cells as EPC. Endothelial progenitor cell proliferation by vascular endothelial growth factor was evaluated by the tetrazolium method. Endothelial progenitor cell migration in response to stromal-derived factor-1α was also evaluated by using a Boyden chamber assay. Results: The percentage of cells positively stained by direct fluorescent staining by acetylated low-density lipoprotein and isolectin B4 was similar between fresh and Cryo-EPC (EPC = 96.0 ± 0.42 versus Cryo-EPC = 95.2 ± 1.2; p = 0.21). Vascular endothelial growth factor increased proliferation activity in fresh and Cryo-EPC (p < 0.01). Stromal-derived factor-1α increased migration activity in fresh and Cryo-EPC (p < 0.01). There was no difference in proliferation and migration activity between fresh and Cryo-EPC. Conclusions: Ex vivo expansion by cell culture was a useful method for collection of bone marrow-derived EPC from cryopreserved mononuclear cells. Proliferation and migration function of EPC is preserved after cryopreservation.
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U2 - 10.1016/j.athoracsur.2007.12.006
DO - 10.1016/j.athoracsur.2007.12.006
M3 - Article
C2 - 18355528
AN - SCOPUS:40749129948
VL - 85
SP - 1361
EP - 1366
JO - Annals of Thoracic Surgery
JF - Annals of Thoracic Surgery
SN - 0003-4975
IS - 4
ER -