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Statements

Subject Item
n2:RIV%2F67985904%3A_____%2F00%3A21003028%21RIV%2F2003%2FAV0%2FA21003%2FN
rdf:type
n7:Vysledek skos:Concept
dcterms:description
The success of somatic nuclear transfer critically depends on the cell cycle stage of the donor nucleus and the recipient cytoplast. In this study we tested serum deprivation as well as two reversible cell cycle inhibitors, aphidicolin and butyrolactone II, for their ability to synchronize porcine fetal fibroblasts at either G0 stage or G1/S or G2/M transition. The synchronization efficiency of the various protocols was determined by fluorescence-activated cell sorting (FACS), cell proliferation assays, and semiquantitative multiplex reverse transcription-polymerase chain reaction detection of the cell cycle-regulated porcine Polo-like kinase mRNA (Plk-p). FAGS measurements revealed that 66.6-73.3% of the porcine fetal fibroblasts were in G0/G1 stage (2C DNA content) in serum-supplemented medium. Short periods of 24-72 h of serum deprivation significantly increased the proportion of cells at G0/G1 phase to 77.9-80.2%, and mitotic activity had already terminated after 48 h. Prolonged culture in serum-d The success of somatic nuclear transfer critically depends on the cell cycle stage of the donor nucleus and the recipient cytoplast. In this study we tested serum deprivation as well as two reversible cell cycle inhibitors, aphidicolin and butyrolactone II, for their ability to synchronize porcine fetal fibroblasts at either G0 stage or G1/S or G2/M transition. The synchronization efficiency of the various protocols was determined by fluorescence-activated cell sorting (FACS), cell proliferation assays, and semiquantitative multiplex reverse transcription-polymerase chain reaction detection of the cell cycle-regulated porcine Polo-like kinase mRNA (Plk-p). FAGS measurements revealed that 66.6-73.3% of the porcine fetal fibroblasts were in G0/G1 stage (2C DNA content) in serum-supplemented medium. Short periods of 24-72 h of serum deprivation significantly increased the proportion of cells at G0/G1 phase to 77.9-80.2%, and mitotic activity had already terminated after 48 h. Prolonged culture in serum-d
dcterms:title
Cell cycle synchronization of porcine fetal fibroblasts: Effects of serum deprivation and reversible cell cycle inhibitors. Cell cycle synchronization of porcine fetal fibroblasts: Effects of serum deprivation and reversible cell cycle inhibitors.
skos:prefLabel
Cell cycle synchronization of porcine fetal fibroblasts: Effects of serum deprivation and reversible cell cycle inhibitors. Cell cycle synchronization of porcine fetal fibroblasts: Effects of serum deprivation and reversible cell cycle inhibitors.
skos:notation
RIV/67985904:_____/00:21003028!RIV/2003/AV0/A21003/N
n4:strany
412;419
n4:aktivita
n9:P n9:Z
n4:aktivity
P(KSK2038602), Z(AV0Z5045916)
n4:cisloPeriodika
2
n4:dodaniDat
n12:2003
n4:domaciTvurceVysledku
n5:2188554 n5:5434505
n4:druhVysledku
n10:J
n4:duvernostUdaju
n13:S
n4:entitaPredkladatele
n14:predkladatel
n4:idSjednocenehoVysledku
706218
n4:idVysledku
RIV/67985904:_____/00:21003028
n4:jazykVysledku
n18:eng
n4:klicovaSlova
N/A
n4:klicoveSlovo
n16:N%2FA
n4:kodStatuVydavatele
US - Spojené státy americké
n4:kontrolniKodProRIV
[652B7E4EF059]
n4:nazevZdroje
Biology of Reproduction
n4:obor
n17:EB
n4:pocetDomacichTvurcuVysledku
2
n4:pocetTvurcuVysledku
6
n4:pocetUcastnikuAkce
0
n4:pocetZahranicnichUcastnikuAkce
0
n4:projekt
n15:KSK2038602
n4:rokUplatneniVysledku
n12:2000
n4:svazekPeriodika
62
n4:tvurceVysledku
Motlík, Jan Niemann, H. Paul, D. Carnwath, J. W. Kues, W. A. Anger, Martin
n4:zamer
n11:AV0Z5045916
s:issn
0006-3363
s:numberOfPages
8