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Three-dimensional Myoblast Aggregates--Effects of Modeled MicrogravityThe overall objective of these studies is to elucidate the molecular and cellular alterations that contribute to muscle atrophy in astronauts caused by exposure to microgravity conditions in space. To accomplish this, a three-dimensional model test system was developed using mouse myoblast cells (C2C12). Myoblast cells were grown as three-dimensional aggregates (without scaffolding or other solid support structures) in both modeled microgravity (Rotary Cell Culture System, Synthecon, Inc.) and at unit gravity in coated Petri dishes. Evaluation of H&E stained thin sections of the aggregates revealed the absence of any necrosis. Confocal microscopy evaluations of cells stained with the Live/Dead assay (Molecular Probes) confirmed that viable cells were present throughout the aggregates with an average of only three dead cells observed per aggregate. Preliminary results from gene array analysis (Affymetrix chip U74Av2) showed that approximately 14% of the genes were down regulated (decreased more than 3 fold) and 4% were upregulated in cells exposed to modeled microgravity for 12 hours compared to unit gravity controls. Additional studies using fluorescent phallacidin revealed a decrease in F-actin in the cells exposed to modeled microgravity compared to unit gravity. Myoblast cells grown as aggregates in modeled microgravity exhibited spontaneous differentiation into syncitia while no differentiation was seen in the unit gravity controls. These studies show that 1)the model test system developed is suitable for assessing cellular and molecular alterations in myoblasts; 2) gene expression alterations occur rapidly (within 12 hours) following exposure to modeled microgravity; and 3) modeled microgravity conditions stimulated myoblast cell differentiation. Achieving a greater understanding of the molecular alterations leading to muscle atrophy will eventually enable the development of cell-based countermeasures, which may be valuable for treatment of muscle diseases on Earth and future space explorations.
Document ID
20060010411
Acquisition Source
Johnson Space Center
Document Type
Conference Paper
Authors
Byerly, Diane
(NASA Johnson Space Center Houston, TX, United States)
Sognier, M. A.
(Universities Space Research Association Houston, TX, United States)
Marquette, M. L.
(Texas Univ. Galveston, TX, United States)
Date Acquired
August 23, 2013
Publication Date
January 1, 2006
Subject Category
Aerospace Medicine
Meeting Information
Meeting: American Society of Cell Biology
Location: Washington, DC
Country: United States
Start Date: December 4, 2004
End Date: December 12, 2004
Sponsors: American Society of Cell Biology
Distribution Limits
Public
Copyright
Other

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