CXCR7 is induced by hypoxia and mediates glioma cell migration
ALTERATIONS OF HYPOXIA INDUCIBLE FACTOR −1α IN ENCAPSULATED AND NON–ENCAPSULATED MESENCHYMAL...
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E1823 JACC March 12, 2013 Volume 61, Issue 10 TCT@ACC-i2: Invasive and Interventional Cardiology ALTERATIONS OF HYPOXIA INDUCIBLE FACTOR -1 α IN ENCAPSULATED AND NON-ENCAPSULATED MESENCHYMAL STEM CELLS SUBJECTED TO HYPOXIA Poster Contributions Poster Sessions, Expo North Sunday, March 10, 2013, 3:45 p.m.-4:30 p.m. Session Title: Cell Therapy and Angiogenesis Abstract Category: 42. TCT@ACC-i2: Cell Therapy & Angiogenesis Presentation Number: 2110-235 Authors: Dorota Anna Kedziorek, Yingli Fu, Yi Xu, Kathleen Gabrielson, Dara Kraitchman, Johns Hopkins University, Baltimore, MD, USA Background: Previously, we have shown that microencapsulated allogeneic mesenchymal stem cells (MSCs) improve angiogenesis and arteriogenesis compared to non-encapsulated MSCs in a rabbit model of peripheral arterial disease1. Whether this improvement is related to enhanced cell survival due to immunoprotection or enhanced ability of the MSCs to release pro-angiogenic cytokines in response to hypoxia is not clear. The objective of the in vitro current study was to evaluate the response to hypoxia of microencapsulated and non-encapsulated human MSCs (hMSCs). Methods: Early passage hMSCs were microencapsulated in a 2% alginate solution or perfluorocarbon-modified alginate (PFOB) containing 106 hMSCs /ml alginate. Encapsulated and non-encapsulated MSCs were subjected to 1% oxygen conditions for 4 hours and compared to controls in normoxic conditions (5% CO2). Alginate and PFOB capsules were lysed with citric acid and Versene and proteins were isolated. Western immunoblots were probed with anti-human hypoxia inducible factor-1α (HIF-1α) monoclonal antibody. Results: Immunoblot analysis of cell lysates revealed increased level of HIF-1α expression in all experimental arms cultured in hypoxia as compared to control, with a trend towards less HIF-1α in encapsulated cells relative to cancer or naked cells (P<0.07). PFOB encapsulated hMSCs demonstrated production of a small amount of HIF-1α when cultured in normoxia (0.09±0.06) and hypoxia (0.02±0.01). Discussion: MSCs showed evidence of less hypoxic protein upregulation when they are encapsulated, suggesting that encapsulation may be the most beneficial for angiogenic recruitment in PAD therapy. References: (1) Kedziorek et al. Stem Cells, 2012.
Transcript of ALTERATIONS OF HYPOXIA INDUCIBLE FACTOR −1α IN ENCAPSULATED AND NON–ENCAPSULATED MESENCHYMAL...
E1823JACC March 12, 2013Volume 61, Issue 10
TCT@ACC-i2: Invasive and Interventional CardiologyalTeraTions of hypoxia induciBle facTor -1α in encapsulaTed and non-encapsulaTed mesenchymal sTem cells suBjecTed To hypoxia
Poster ContributionsPoster Sessions, Expo NorthSunday, March 10, 2013, 3:45 p.m.-4:30 p.m.
Session Title: Cell Therapy and AngiogenesisAbstract Category: 42. TCT@ACC-i2: Cell Therapy & AngiogenesisPresentation Number: 2110-235
Authors: Dorota Anna Kedziorek, Yingli Fu, Yi Xu, Kathleen Gabrielson, Dara Kraitchman, Johns Hopkins University, Baltimore, MD, USA
Background: Previously, we have shown that microencapsulated allogeneic mesenchymal stem cells (MSCs) improve angiogenesis and arteriogenesis compared to non-encapsulated MSCs in a rabbit model of peripheral arterial disease1. Whether this improvement is related to enhanced cell survival due to immunoprotection or enhanced ability of the MSCs to release pro-angiogenic cytokines in response to hypoxia is not clear. The objective of the in vitro current study was to evaluate the response to hypoxia of microencapsulated and non-encapsulated human MSCs (hMSCs).
methods: Early passage hMSCs were microencapsulated in a 2% alginate solution or perfluorocarbon-modified alginate (PFOB) containing 106 hMSCs /ml alginate. Encapsulated and non-encapsulated MSCs were subjected to 1% oxygen conditions for 4 hours and compared to controls in normoxic conditions (5% CO2). Alginate and PFOB capsules were lysed with citric acid and Versene and proteins were isolated. Western immunoblots were probed with anti-human hypoxia inducible factor-1α (HIF-1α) monoclonal antibody.
results: Immunoblot analysis of cell lysates revealed increased level of HIF-1α expression in all experimental arms cultured in hypoxia as compared to control, with a trend towards less HIF-1α in encapsulated cells relative to cancer or naked cells (P<0.07). PFOB encapsulated hMSCs demonstrated production of a small amount of HIF-1α when cultured in normoxia (0.09±0.06) and hypoxia (0.02±0.01).
discussion: MSCs showed evidence of less hypoxic protein upregulation when they are encapsulated, suggesting that encapsulation may be the most beneficial for angiogenic recruitment in PAD therapy.
references: (1) Kedziorek et al. Stem Cells, 2012.
