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Cryopreservation of mouse morulae through different methods: slow-freezing, vitrification and quick-freezing Braz. J. Vet. Res. Anim. Sci.
MELLO,Marco Roberto Bourg de; QUEIROZ,Vinícius Seixas; LIMA,Alessandra Sobreira de; TAVARES,Liliam Mara Trevisam; ASSUMPÇÃO,Mayra Elena Ortiz D'Avila; WHEELER,Mathew B.; VISINTIN,José Antonio.
The in vitro and in vivo development of mouse morulae after cryopreservation through different methods was examined. The slow-freezing involved an equilibration in 1.5M ethylene glycol (EG) and cooled at 0.5; 0.7; 1.0 or 1.2ºC/minute. The vitrification involved a 3 minutes equilibration in 20% EG and 60 seconds in solution containing 40% EG, 18% ficoll and 10.26% sucrose. The quick-freezing involved an equilibration in 3M EG + 0.3M sucrose for 5 minutes and 2 minutes in nitrogen vapor. In all three methods the straws were thawed in air for 10 seconds and in water at 25ºC for 20 seconds and the embryos cultured in vitro for 72 hours to estimate blastocyst rate. To assess viability in vivo, frozen morulae as well as fresh embryos were transferred into...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Cryopreservation; Embryo; Mouse.
Ano: 2001 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-95962001000400003
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Effects of cryoprotectant and plunging temperature in liquid nitrogen on the in vitro and in vivo development of murine morulae Braz. J. Vet. Res. Anim. Sci.
PANTANO,Thaís; MELLO,Marco Roberto Bourg de; GARCIA,José Fernando; HO,Linda Lee; VISINTIN,José Antonio.
The effects of plunging temperature in liquid nitrogen and cryoprotectant dilution methods were evaluated for compacted mouse morulae frozen in 1.5 M ethylene-glycol (E), 1.5M propylene-glycol (P) or 1.4M glycerol (G). Morulae were equilibrated for 10 minutes in cryoprotectant solution and loaded into 0.25 ml straws with cryoprotectant solution in 3 columns (groups E1, P1, G1) or cryoprotectant in the center and PBS in the lateral columns (E2, P2). Straws were cooled at 0.5ºC/min to -25 or -30ºC and plunged into liquid nitrogen. Straws were thawed in water at 22ºC for 20 seconds. Cryoprotectant was diluted in 3 steps for group G1 and in one step for groups E1 and P1 (direct transfer to PBS + 10% FCS) and E2 and P2 (shaken to mix the 3 columns before...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Embryos; Cryopreservation; Glycerol; Ethylene glycol; Propanediols.
Ano: 2000 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-95962000000300012
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