Bio-scaffold Versus Synthetic Scaffold Interactions With Seeded Stem Cells In Dynamic Flow Culture Environments
Chia-Pei Denise Hsu, Brittany A. Gonzalez, Sharan Ramaswamy.
Florida International University, Miami, FL, USA.
OBJECTIVE: Elastin is an important component for extracellular matrix and cardiac tissue regeneration (1). The objective of this study is to determine whether porcine small intestinal submucosa (PSIS) bio-scaffolds can better promote cardiac tissue regeneration from bone marrow stem cells compared to poly glycolic acid-poly L-lactic acid (PGA-PLLA) synthetic scaffolds under physiologically-relevant flow environments.
METHODS: Human bone marrow stem cells (RoosterBio, Frederick, MD) were seeded at 2,000,000 cells per 2.5cm2 in 1.5cm x 1cm PSIS bio-scaffolds and PGA-PLLA synthetic scaffolds using Dulbecco's Modified Eagle Medium (DMEM) at 10% fetal bovine serum (FBS), 1% penicillin-streptomycin (P/S), 82ug/mL L-ascorbic acid 2-phosphate (AA2P), and 2ng/mL basic fibroblast growth factor (bFGF). The seeded PSIS strips were placed in rotisserie for 8 days followed by conditioning in a bioreactor under physiologically-relevant flow environment with surface shear stress of 3dyne/cm2 for an additional 14 days. Conditioned samples were then fixed in 10% formalin at 4degC overnight, embedded in optimal cutting temperature (OCT) compound, and sectioned at 16 um using a cryostat. Sectioned samples were subsequently stained with elastin mouse monoclonal primary antibody (Novus Biologicals, Littleton, CO), followed by goat anti-mouse polyclonal secondary antibody (Thermo Fisher, Waltham, MA) and 4',6-diamidino-2phenylindole (DAPI). Stained samples were imaged using a confocal microscope.
RESULTS: Image results show presence of elastin protein (green) in the PSIS strip (a) and none in the PGA-PLLA (b) samples.
CONCLUSIONS: Cells seeded in PSIS bio-scaffold extracellular matrices facilitate higher production of elastin under flow mechanical conditions compared to PGA-PLLA synthetic scaffolds. Bio-scaffolds extracellular components allow bone marrow stem cells to communicate and secrete engineered matrix component, such as elastin that will be useful for enhancing cardiovascular regeneration.
REFERENCE(S): (1) Gonzalez de Torre, I. et al: Frontiers in Bioengineering and Biotechnology 2020-Jun-30; Vol. 8.
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