|
KMID : 0900919990230020141
|
|
Korean journal of Animal Reproduction
1999 Volume.23 No. 2 p.141 ~ p.148
|
|
|
Cryopreservation of Bovine IVM/IVF/IVC Hatched Blastocysts
|
|
Lee K.-S.
Kim E.-Y.
Yi B.-K.
Nam H.-K.
Yoon S.-H.
Park S.-P.
Lim J.-H.
|
|
|
Abstract
|
|
|
|
This study was to test whether the viability of bovine hatched blastocysts (HBs) can be maintained after vitrification and thawing. The HBs were produced in vitro at Day 9 and Day 10 after IVF, and they were classified to small (S-HBs; øleq300 {mu}{textrm}{m}) and large(L-HBs; ø＞300 {mu}{textrm}{m}) on the basis of embryo diameter using eyepiece micrometer. As freezing solution, we used EFS35 which consisted of 35% ethylene glycol (EG), 18% ficoll, 0.3 M sucrose and 10% FBS added in mDPBS. Vitrification was taken by two-step method, the HBs were equilibrated in 10% EG for 5 minutes and then shortly exposed in EFS35 and plunged into LN_2for 30~45 sec. After thawing, the survival rates were assessed by the re-expansion of the blastocoel during 2 h and 16 h of culture. The results obtained in these experiments were summarized as follows; 1) When the blastocysts(40.8%) recovered at Day 8 after IVF were further cultured for 24 h(Day 9 after IVF) and 48 h(Day 10 after IVF), the rates of HBs were 20.5% and 6.7%, respectively. Also, the total cell number of HBs on Day 9 was significantly higher than that of HBs on Day 10 (p＜0.01). 2) When the effects of freezing solution to the survival of Day 9 L-HBs were examined, the rate of vitrified group (75.7%) was significantly lower than 100% of control and exposed group(p＜0.05). 3) When the survival rates of vitrified HBs according to size and developmental age were examined, the data of L-HBs (75.5%) and S-HBs(63.6%) on Day 9 were slightly higher than those of L-HBs(64.3%) and S-HBs(60.7%) on Day 10. 4) Also, when the in vitro survival of Day 9 HBs was evaluated under different culture condition after thawing, the result in culture medium only (79.3%) was significantly higher than 43.2% in co-culture group (p＜0.05). These results demonstrated that bovine HBs can be successfully cryopreserved by two-step vitrification method using EFS35.
|
|
|
KEYWORD
|
|
|
Hatched blastocyst, Vitrification, EFS35, In vitro survival
|
|
|
FullTexts / Linksout information
|
|
|
|
|
Listed journal information
|
|
|
|
|