Nedambale T L, Du F, Xu J, Chaubal S A, Dinnyes A, Groen W, Faber D, Dobrinsky J R, Yang X, Tian X C
Center for Regenerative Biology/Department of Animal Science, University of Connecticut, 1392 Storrs Rd., U-4243, Storrs, CT 06269, USA.
Theriogenology. 2006 Nov;66(8):1951-60. doi: 10.1016/j.theriogenology.2006.04.044. Epub 2006 Jun 19.
This study was conducted to compare the efficacy of four in vitro fertilization (IVF) media: Bracket and Oliphant's medium (BO), modified medium 199 (IVF-M199), modified Tyrode's medium (MTM), and modified KSOM (m-KSOM) on fertilization efficiency and blastocyst formation rate. In addition, we wanted to investigate the benefit of prolonging the IVF period (from 6 to 18 h) using the two most effective IVF media determined in our initial experiment; subsequently, blastocyst viability was assessed following vitrification. A higher incidence of polyspermic fertilization was observed in the MTM (6%) and in BO, in both the 6 and 18 h (7% and 11%, respectively) groups, than in the m-KSOM (1%) or in the IVF-M199 6 or 18 h (1 and 3%, respectively) groups. Cleavage rates were similar in BO, IVF-M199, and MTM 48 h post-fertilization; however, the lowest cleavage rate was observed for m-KSOM. A greater proportion of zygotes developed into 8-cell embryos in IVF-M199 than in other IVF media. Subsequently, a greater proportion of blastocyst formation and hatching was achieved in IVF-M199 (40% and 79%, respectively) or BO (35% and 74%, respectively) than in m-KSOM (18% and 58%, respectively) or MTM (22% and 66%, respectively). Prolonging IVF to 18 h did not alter cleavage rates; however, the highest rate of overall blastocyst formation was achieved in the IVF-M199 18 h (49%), rather than in the BO 18 h (20%) group. Vitrified/thawed blastocysts from IVF-M199 groups re-expanded and developed better, as compared to the BO 18 h group, and hatching rate and total cell number in IVF-M199 18 h group was comparable to the control groups (non-vitrified). Vitrification reduced survival compared to controls. In conclusion, IVF-M199 was successfully used for IVF, compared favorably to BO medium, and offered the advantage of an extended IVF period for up to 18 h that requires only one-half a dose of semen, and resulted in better quality blastocysts that endured vitrification with a hatching rate comparable to that of control groups.
本研究旨在比较四种体外受精（IVF）培养基：布雷克特和奥利芬特培养基（BO）、改良培养基199（IVF-M199）、改良泰勒氏培养基（MTM）和改良的钾离子简单培养基（m-KSOM）对受精效率和囊胚形成率的影响。此外，我们想研究使用在我们初步实验中确定的两种最有效的IVF培养基将IVF时间延长（从6小时延长至18小时）的益处；随后，评估玻璃化后囊胚的活力。与m-KSOM组（1%）或IVF-M199 6小时或18小时组（分别为1%和3%）相比，在MTM组（6%）以及6小时和18小时的BO组（分别为7%和11%）中观察到多精受精的发生率更高。受精后48小时，BO、IVF-M199和MTM的卵裂率相似；然而，m-KSOM的卵裂率最低。与其他IVF培养基相比，IVF-M199中有更大比例的受精卵发育成8细胞胚胎。随后，与m-KSOM组（分别为18%和58%）或MTM组（分别为22%和66%）相比，IVF-M199组（分别为40%和79%）或BO组（分别为35%和74%）实现了更高比例的囊胚形成和孵化。将IVF延长至18小时并没有改变卵裂率；然而，IVF-M199 18小时组（49%）的总体囊胚形成率最高，而不是BO 18小时组（20%）。与BO 18小时组相比，IVF-M199组的玻璃化/解冻囊胚重新扩张且发育更好，IVF-M199 18小时组的孵化率和总细胞数与对照组（未玻璃化）相当。与对照组相比，玻璃化降低了存活率。总之，IVF-M199成功用于IVF，与BO培养基相比具有优势，可将IVF时间延长至18小时，且只需要一半剂量的精液，并产生质量更好的囊胚，这些囊胚能够耐受玻璃化，孵化率与对照组相当。
