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Quantifying the Effects of Brain-Death on Porcine Muscle-Derived Stem Cells: An In vitro Study
Anil Chaturvedi, Samuel Boas, Corinne Wee, James Reynolds, Anand Kumar
Case Western Reserve University
2019-02-15
Presenter: Anil Chaturvedi
Affidavit:
I certify that the material proposed for presentation in this abstract has not been published in any scientific journal or previously presented at a major meeting. This project represents the original work of the student.
Director Name: Anand Kumar
Author Category: Medical Student
Presentation Category: Basic Science Research
Abstract Category: General Reconstruction
Background: Muscle-derived stem cells (MDSCs) are multipotent cells with both immuno-regulatory and regenerative capacity. We hypothesize that brain-death negatively affects the stem cell-niche in vascularized composite allografts (VCA) and may affect tolerance/rejection. Our study aim was to evaluate the in vitro properties of MDSCs in a porcine brain-death VCA model.
Methods: MDSCs were isolated from porcine hind limbs pre- and post-brain-death using a collagen-sorting preplate technique. Imaging was obtained at various time points (t=0-10 days). In vitro measurements were time to confluence, migration, cell size, and cell density. Images were analyzed using ImageJ, statistical analysis was performed using Mathematica.
Results: Time to confluence was significantly different, (t=3, 5, 7, 10 days) pre vs. post brain death were 20%, 38%, 75%, and 81% vs. 7%, 17%, 31%, 39% (p<0.01). Migration was not significantly different, (t=48 hours) pre vs. post-brain-death (88% vs 81%) (p=0.10). Cell size was not significantly different, (t=10 days) pre vs. post-brain-death were 11µm vs. 9µm (p=0.3). Cell density was not significantly different, (t=10 days) pre vs. post-brain-death were 154,000 cells/ml vs. 114,000 cells/ml (p=0.6).
Conclusion: Brain-death significantly impairs MDSC expansion and mitosis based on time to confluence. Brain-death does not appear to impact MDSC cellular migration, size and cell density at ultimate confluence. Further studies will investigate the effects of brain-death on MDSC tri-lineage differentiation, myotube formation, and immune modulation.
