Collagen often acts as an extracellular and intracellular marker for in vitro experiments, and its quality defines tissue constructs. To validate collagen detection techniques, cardiac valve interstitial cells were isolated from pigs and cultured under two different conditions; with and without ascorbic acid. The culture with ascorbic acid reached higher cell growth and collagen deposition, although the expression levels of collagen gene stayed similar to the culture without ascorbic acid. The fluorescent microscopy was positive for collagen fibers in both the cultures. Visualization of only extracellular collagen returned a higher correlation coefficient when comparing the immunolabeling and second harmonic generation microscopy images in the culture with ascorbic acid. Lastly, it was proved that the hydroxyproline strongly contributes to the second-order susceptibility tensor of collagen molecules, and therefore the second harmonic generation signal is impaired in the culture without ascorbic acid.
- MeSH
- barvení a značení MeSH
- buněčné kultury MeSH
- kolagen typu I analýza genetika metabolismus MeSH
- kultivované buňky MeSH
- Leydigovy buňky chemie metabolismus MeSH
- prasata MeSH
- srdeční chlopně chemie cytologie metabolismus MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- MeSH
- alciánová modř diagnostické užití MeSH
- autoradiografie metody využití MeSH
- barvení a značení metody využití MeSH
- glykosaminoglykany chemie metabolismus MeSH
- histocytochemie metody využití MeSH
- kalcinóza diagnóza metabolismus MeSH
- kardiomyocyty metabolismus patologie MeSH
- králíci MeSH
- sírany diagnostické užití chemie MeSH
- srdeční chlopně chemie metabolismus MeSH
- statistika jako téma MeSH
- toloniumchlorid diagnostické užití MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- zvířata MeSH
