Document Detail


An in vitro system to evaluate the effects of ischemia on survival of cells used for cell therapy.
MedLine Citation:
PMID:  17417737     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Maintaining cell viability is a major challenge associated with transplanting cells into ischemic myocardium to restore function. A likely contributor to significant cell death during cardiac cell therapy is hypoxia/anoxia. We developed a system that enabled quantification and association of cell survival with oxygen and nutrient values within in vitro constructs. Myoblasts were suspended in 2% collagen gels in 1 cm diameter x 1 cm deep constructs. At 48 +/- 3 h post-seeding, oxygen levels were measured using microelectrodes and gels were snap-frozen. Bioluminescence metabolite imaging and TUNEL staining were performed on cryosections. Oxygen and glucose consumption and lactate production rates were calculated by fitting data to Fick's second law of diffusion with Michaelis-Menten kinetics. Oxygen levels dropped to 0 mmHg and glucose levels dropped from 4.28 to 3.18 mM within the first 2000 mum of construct depth. Cell viability dropped to approximately 40% over that same distance and continued to drop further into the construct. We believe this system provides a reproducible and controllable test bed to compare survival, proliferation, and phenotype of various cell inputs (e.g., myoblasts, mesenchymal stem cells, and cardiac stem cells) and the impact of different treatment regimens on the likelihood of survival of transplanted cells.
Authors:
Bryce H Davis; Thies Schroeder; Pavel S Yarmolenko; Farshid Guilak; Mark W Dewhirst; Doris A Taylor
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Publication Detail:
Type:  In Vitro; Journal Article; Research Support, N.I.H., Extramural     Date:  2007-04-07
Journal Detail:
Title:  Annals of biomedical engineering     Volume:  35     ISSN:  0090-6964     ISO Abbreviation:  Ann Biomed Eng     Publication Date:  2007 Aug 
Date Detail:
Created Date:  2007-07-16     Completed Date:  2007-09-26     Revised Date:  2007-12-03    
Medline Journal Info:
Nlm Unique ID:  0361512     Medline TA:  Ann Biomed Eng     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1414-24     Citation Subset:  IM    
Affiliation:
Department of Biomedical Engineering, Duke University, Durham, NC, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Anoxia
Cell Survival
Collagen Type I / metabolism
Glucose / metabolism*
Lactic Acid / metabolism
Mesenchymal Stem Cells / physiology
Muscle, Skeletal / cytology,  physiology*
Myoblasts, Cardiac / physiology*,  transplantation
Myocardial Ischemia*
Oxygen Consumption / physiology*
Swine
Tissue Transplantation
Grant Support
ID/Acronym/Agency:
HL-63703/HL/NHLBI NIH HHS; R-01 HL-63346/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
0/Collagen Type I; 50-21-5/Lactic Acid; 50-99-7/Glucose

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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