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Statements

Subject Item
n2:RIV%2F60076658%3A12520%2F14%3A43886839%21RIV15-MSM-12520___
rdf:type
skos:Concept n19:Vysledek
rdfs:seeAlso
http://www.agriculturejournals.cz/publishedArticles/CJAS/2014-59-8-381.pdf
dcterms:description
A practical technique for isolation and cryopreservation of tench (Tinca tinca) (Cyprinidae, Teleostei) early stages of germ cells (GC), including spermatogonia and spermatocytes, is reported for the first time. The germ-line cells possess the ability to differentiate into functional gametes of both sexes. These early stages of germ cells are small enough to be well-suited to cryopreservation, which, together with their high level of plasticity, makes their preservation a promising tool for maintaining genetic resources. Testicular cells were distinguished and separated by Percoll gradient, with the highest proportion of GC (62.2%) obtained from the 30% layer. The concentration and viability of GC were determined, and specific staining (DDX4) for germ cells was used to distinguish GC from somatic cells. Early stages of germ cells were cryopreserved in an extender composed of phosphate buffered saline (pH 8) with 0.5% BSA, 50mM D-glucose, and containing 1.5M cryoprotectant in the pre-programmed PLANER Kryo10 series III using a cooling protocol from +10 degrees C to -80 degrees C at a rate of 1 degrees C/min. The effect of six cryoprotectants - methanol, dimethyl sulfoxide, dimethyl sulfoxide + propanediol (1 : 1), glycerol, ethylene glycol, and dimethylacetamid was assessed, and the results were evaluated by comparing the percentage of viable frozen/thawed GC by ANOVA, Tukey's HSD test (P { 0.05). Almost the same viability rates were obtained with no significant differences among tested cryoprotectants, indicating high stability of GC in cryoprotectants. Nevertheless, glycerol at a concentration of 1.5M was associated with the highest survival rate of thawed tench GC (57.69 +/- 16.85%). A practical technique for isolation and cryopreservation of tench (Tinca tinca) (Cyprinidae, Teleostei) early stages of germ cells (GC), including spermatogonia and spermatocytes, is reported for the first time. The germ-line cells possess the ability to differentiate into functional gametes of both sexes. These early stages of germ cells are small enough to be well-suited to cryopreservation, which, together with their high level of plasticity, makes their preservation a promising tool for maintaining genetic resources. Testicular cells were distinguished and separated by Percoll gradient, with the highest proportion of GC (62.2%) obtained from the 30% layer. The concentration and viability of GC were determined, and specific staining (DDX4) for germ cells was used to distinguish GC from somatic cells. Early stages of germ cells were cryopreserved in an extender composed of phosphate buffered saline (pH 8) with 0.5% BSA, 50mM D-glucose, and containing 1.5M cryoprotectant in the pre-programmed PLANER Kryo10 series III using a cooling protocol from +10 degrees C to -80 degrees C at a rate of 1 degrees C/min. The effect of six cryoprotectants - methanol, dimethyl sulfoxide, dimethyl sulfoxide + propanediol (1 : 1), glycerol, ethylene glycol, and dimethylacetamid was assessed, and the results were evaluated by comparing the percentage of viable frozen/thawed GC by ANOVA, Tukey's HSD test (P { 0.05). Almost the same viability rates were obtained with no significant differences among tested cryoprotectants, indicating high stability of GC in cryoprotectants. Nevertheless, glycerol at a concentration of 1.5M was associated with the highest survival rate of thawed tench GC (57.69 +/- 16.85%).
dcterms:title
Isolation and cryopreservation of early stages of germ cells of tench (Tinca tinca) Isolation and cryopreservation of early stages of germ cells of tench (Tinca tinca)
skos:prefLabel
Isolation and cryopreservation of early stages of germ cells of tench (Tinca tinca) Isolation and cryopreservation of early stages of germ cells of tench (Tinca tinca)
skos:notation
RIV/60076658:12520/14:43886839!RIV15-MSM-12520___
n3:aktivita
n11:S n11:P
n3:aktivity
P(ED2.1.00/01.0024), P(LH13246), P(LO1205), S
n3:cisloPeriodika
8
n3:dodaniDat
n10:2015
n3:domaciTvurceVysledku
Saito, Taiju n8:9895744 n8:7078935 n8:4388259 n8:5876001 n8:5744423
n3:druhVysledku
n15:J
n3:duvernostUdaju
n7:S
n3:entitaPredkladatele
n4:predkladatel
n3:idSjednocenehoVysledku
22953
n3:idVysledku
RIV/60076658:12520/14:43886839
n3:jazykVysledku
n18:eng
n3:klicovaSlova
viability; cryoprotectant; Percoll gradient; transplantation; isolation; germ cells
n3:klicoveSlovo
n6:cryoprotectant n6:transplantation n6:viability n6:germ%20cells n6:Percoll%20gradient n6:isolation
n3:kodStatuVydavatele
CZ - Česká republika
n3:kontrolniKodProRIV
[13968A5CC23E]
n3:nazevZdroje
Czech Journal of Animal Science : Živočišná výroba
n3:obor
n16:EB
n3:pocetDomacichTvurcuVysledku
6
n3:pocetTvurcuVysledku
6
n3:projekt
n13:LO1205 n13:LH13246 n13:ED2.1.00%2F01.0024
n3:rokUplatneniVysledku
n10:2014
n3:svazekPeriodika
59
n3:tvurceVysledku
Gazo, Ievgenia Linhartová, Zuzana Güralp, Hilal Rodina, Marek Saito, Taiju Pšenička, Martin
n3:wos
000342060900005
s:issn
1212-1819
s:numberOfPages
10
n14:organizacniJednotka
12520