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Valve interstitial cell culture: Production of mature type I collagen and precise detection
J. Liskova, D. Hadraba, E. Filova, M. Konarik, J. Pirk, K. Jelen, L. Bacakova,
Jazyk angličtina Země Spojené státy americké
Typ dokumentu časopisecké články
Grantová podpora
NV15-29153A
MZ0
CEP - Centrální evidence projektů
Digitální knihovna NLK
Plný text - Článek
Odkazy
PubMed
28455837
DOI
10.1002/jemt.22886
Knihovny.cz E-zdroje
- 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
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.
Faculty of Physical Education and Sport Charles University Prague 162 00 Czech Republic
Institute for Clinical and Experimental Medicine Prague 140 21 Czech Republic
Institute of Physiology the Czech Academy of Sciences Prague 142 20 Czech Republic
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- $a 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.
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