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Establishment of glass catfish (Kryptopterus bicirrhis) fin-derived cells.

Han JE, Choresca CH, Koo OJ, Oh HJ, Hong SG, Kim JH, Shin SP, Jun JW, Lee BC, Park SC - Cell Biol Int Rep (2010) (2011)

Bottom Line: The cells grew well in DMEM (Dulbecco's modified Eagle's medium) containing 1% (v/v) P/S (penicillin-streptomycin) and 10% (v/v) fetal bovine serum at 26°C and showed increased cryopreservation efficiency with the slow-freezing method in the presence of 15% dimethyl sulfoxide.In addition, cell cycle analysis was evaluated based on flow cytometric analysis, and culturing to confluence (>85%) was more effective for synchronizing cells at the G(0)/G(1) stages than roscovitine treatment (<75%).The results from testing the cell's viability following cryopreservation and subjecting the cells to cycle analysis can be useful tools for genetic resource management.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Aquatic Animal Medicine.

ABSTRACT
Genetically manipulated transparent animals were already explored in many species for in vivo study of gene expression, transplantation analysis and cancer biology. However, there are no reports about transparent animals as in vitro genetic resources. In the present study, fin-derived cells from glass catfish (Krytopterus bicirrhis), naturally transparent fish with a visible skeleton and internal organs, were isolated after culturing fin explants and characterized using cryopreservation and cell cycle analysis. The cells grew well in DMEM (Dulbecco's modified Eagle's medium) containing 1% (v/v) P/S (penicillin-streptomycin) and 10% (v/v) fetal bovine serum at 26°C and showed increased cryopreservation efficiency with the slow-freezing method in the presence of 15% dimethyl sulfoxide. In addition, cell cycle analysis was evaluated based on flow cytometric analysis, and culturing to confluence (>85%) was more effective for synchronizing cells at the G(0)/G(1) stages than roscovitine treatment (<75%). This is the first report about cell isolation from transparent animals. The results from testing the cell's viability following cryopreservation and subjecting the cells to cycle analysis can be useful tools for genetic resource management.

No MeSH data available.


Related in: MedlinePlus

Typical histograms of DNA content obtained using flow cytometry of glass catfish fin cells at (A) culturing to confluence, (B) 10 μM roscovitine treatment
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Figure 2: Typical histograms of DNA content obtained using flow cytometry of glass catfish fin cells at (A) culturing to confluence, (B) 10 μM roscovitine treatment

Mentions: The relative percentage of the proportions of cells in the G0/G1, S and G2/M stages were calculated (Table 2), and representative histograms for the cell cycle stage distributions of fin cells of glass catfish treated with different culture conditions are shown (Figure 2). In our study, culturing to confluence yielded over 85% of cells arrested at the G0/G1 phase (85.6833±0.9988). Culture to confluence has been known to cause cells to arrest at the G0/G1 phase by contact inhibition and appears to be one of the most widely used methods prior to SCNT (Campbell et al., 2007).


Establishment of glass catfish (Kryptopterus bicirrhis) fin-derived cells.

Han JE, Choresca CH, Koo OJ, Oh HJ, Hong SG, Kim JH, Shin SP, Jun JW, Lee BC, Park SC - Cell Biol Int Rep (2010) (2011)

Typical histograms of DNA content obtained using flow cytometry of glass catfish fin cells at (A) culturing to confluence, (B) 10 μM roscovitine treatment
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3475438&req=5

Figure 2: Typical histograms of DNA content obtained using flow cytometry of glass catfish fin cells at (A) culturing to confluence, (B) 10 μM roscovitine treatment
Mentions: The relative percentage of the proportions of cells in the G0/G1, S and G2/M stages were calculated (Table 2), and representative histograms for the cell cycle stage distributions of fin cells of glass catfish treated with different culture conditions are shown (Figure 2). In our study, culturing to confluence yielded over 85% of cells arrested at the G0/G1 phase (85.6833±0.9988). Culture to confluence has been known to cause cells to arrest at the G0/G1 phase by contact inhibition and appears to be one of the most widely used methods prior to SCNT (Campbell et al., 2007).

Bottom Line: The cells grew well in DMEM (Dulbecco's modified Eagle's medium) containing 1% (v/v) P/S (penicillin-streptomycin) and 10% (v/v) fetal bovine serum at 26°C and showed increased cryopreservation efficiency with the slow-freezing method in the presence of 15% dimethyl sulfoxide.In addition, cell cycle analysis was evaluated based on flow cytometric analysis, and culturing to confluence (>85%) was more effective for synchronizing cells at the G(0)/G(1) stages than roscovitine treatment (<75%).The results from testing the cell's viability following cryopreservation and subjecting the cells to cycle analysis can be useful tools for genetic resource management.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Aquatic Animal Medicine.

ABSTRACT
Genetically manipulated transparent animals were already explored in many species for in vivo study of gene expression, transplantation analysis and cancer biology. However, there are no reports about transparent animals as in vitro genetic resources. In the present study, fin-derived cells from glass catfish (Krytopterus bicirrhis), naturally transparent fish with a visible skeleton and internal organs, were isolated after culturing fin explants and characterized using cryopreservation and cell cycle analysis. The cells grew well in DMEM (Dulbecco's modified Eagle's medium) containing 1% (v/v) P/S (penicillin-streptomycin) and 10% (v/v) fetal bovine serum at 26°C and showed increased cryopreservation efficiency with the slow-freezing method in the presence of 15% dimethyl sulfoxide. In addition, cell cycle analysis was evaluated based on flow cytometric analysis, and culturing to confluence (>85%) was more effective for synchronizing cells at the G(0)/G(1) stages than roscovitine treatment (<75%). This is the first report about cell isolation from transparent animals. The results from testing the cell's viability following cryopreservation and subjecting the cells to cycle analysis can be useful tools for genetic resource management.

No MeSH data available.


Related in: MedlinePlus