Valve interstitial cell culture: Production of mature type I collagen and precise detection

0477592 - FGÚ 2018 RIV US eng J - Journal Article
Lišková, Jana - Hadraba, Daniel - Filová, Elena - Koňařík, M. - Pirk, J. - Jelen, K. - Bačáková, Lucie
Valve interstitial cell culture: Production of mature type I collagen and precise detection.
Microscopy Research and Technique. Roč. 80, č. 8 (2017), s. 936-942. ISSN 1059-910X. E-ISSN 1097-0029
R&D Projects: GA MŠMT(CZ) LM2015062; GA MZd(CZ) NV15-29153A; GA MŠMT(CZ) ED1.1.00/02.0109
Institutional support: RVO:67985823
Keywords : ascorbic acid * cell culture * collagen * fluorescent microscopy * porcine VIC * second harmonic generation
OECD category: Technologies involving the manipulation of cells, tissues, organs or the whole organism (assisted reproduction)
Impact factor: 1.087, year: 2017

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.
Permanent Link: http://hdl.handle.net/11104/0273987