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Hyaluronic acid/collagen (HA/CN) assay for epithelial mesenchymal transformation (EMT) in cardiac valvulogenesis
Author(s) -
Bick Alexander,
DeLaughter Daniel,
Khademhosseini Ali,
Barnett Joey V.
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.754.5
Subject(s) - explant culture , mesenchymal stem cell , hyaluronic acid , chemistry , in vivo , endocardium , microbiology and biotechnology , in vitro , extracellular matrix , atrioventricular canal , andrology , anatomy , biochemistry , biology , pathology , medicine , heart disease
An in vitro assay system was devised for the study EMT in developing cardiac tissue. Explants of isolated chick atrioventricular (AV) canal cushions were placed on hydrated HA/CN gel lattices. Similar to the CN gel assay, cells grew out of the explant from the endocardium and across the gel surface morphologically transitioning from epithelial cells to mesenchymal cells. This process mimics in vivo embryonic heart valvulogenesis. We first optimized the HA/CN gel composition, finding that a 50 mg/mL HA, 1.5 mg/mL CN produced the highest explants viability and were 1000x more mechanically robust than 3D collagen gels with similar endothelial viability. Explants on CN were viable up to 4 days, while HA/CN explants were viable for up to six weeks. We compared the HA/CN to CN gels assay in terms of specificity and gain of function experiments. Though cells began to morphologically undergo EMT on HA/CN gels, they rarely invaded. While further optimization is required before HA/CN can fully recapitulate AV cushion EMT, the enhanced stability and longevity of HA/CN provides a powerful tool to potentially derive novel cardiac cushion endothelial cell lines.