Production of viable bovine blastocysts in defined in vitro conditions.
Production of viable bovine blastocysts in defined in vitro conditions.
复制标题
在规定的体外条件下产生可存活的牛囊胚。
作者：
Levent Keskintepe;Clay A. Burnley;B. Brackett
我们的目的是通过在不含血清和 BSA 的培养基（规定条件）中进行体外成熟、受精和培养 (IVMFC) 来获得可行的囊胚，并记录胚胎移植 (ET) 后妊娠开始时的活力。卵母细胞在含有 100 微克/ml 羊 (o)LH 的改良 TCM 199 (mTCM 199) 中成熟，在基于 TALP 或成分确定培养基 (DM) 的培养基中授精，并在用 6 mg/ml 聚乙烯醇 (PVA) 代替 BSA 制备的合成输卵管液 (SOF) 中培养长达 9 天，并用经过实验修改的 25 mM HEPES 缓冲。胚胎培养的修改包括补充最低必需培养基氨基酸 (MEM)、最低必需培养基非必需氨基酸 (NEA)、MEM 和 NEA 的组合、柠檬酸盐 (c; 0.5 mM)、谷氨酰胺 (1 mM) 或这些的组合。选择用于 IVM 的未成熟卵母细胞中卵裂 (IVF) 和 SOF 中达到囊胚期的比例分别为 66.3% 和 10.9%。在 SOF 中补充柠檬酸盐和非必需氨基酸（即 c-SOF + NEA）可使选择用于 IVM 的卵母细胞的卵裂率达到 85.1%，囊胚发育率达到 42.6%。结合 mTCM 199 中的 IVM 加 100 微克/ml oLH 和 c-SOF + NEA 中的 IVC，从受精培养基中消除蛋白质的努力表明，用 PVA 代替 BSA 制备的改良 DM (mDM) 优于用 PVA 制备的 TALP；囊胚发育的 IVMFC 数据分别为 27.4% 和 18.2% (p < 0.05)。当包含 BSA 或 PVA 时，使用 mDM 进行精子制备和 IVF 产生了可比的囊胚发育。（摘要截断为 250 字）
Our purpose was to obtain viable blastocysts via in vitro maturation, fertilization, and culture (IVMFC) in serum- and BSA-free media (defined conditions) and to document viability by pregnancy initiation following embryo transfer (ET). Oocytes were matured in modified TCM 199 (mTCM 199) with 100 micrograms/ml ovine (o)LH, inseminated in TALP- or defined medium (DM)-based media, and cultured up to 9 days in synthetic oviductal fluid (SOF) prepared with 6 mg/ml polyvinyl alcohol (PVA) instead of BSA and buffered with 25 mM HEPES with experimental modifications. Modifications for embryo culture included supplementation with Minimum Essential Medium amino acids (MEM), Minimum Essential Medium nonessential amino acids (NEA), the combination of MEM and NEA, citrate (c; 0.5 mM), glutamine (1 mM), or combinations of these. Proportions of immature oocytes selected for IVM that cleaved (IVF) and that reached the blastocyst stage in SOF were 66.3% and 10.9%, respectively. Supplementation of SOF with citrate and nonessential amino acids (i.e., c-SOF + NEA) enabled 85.1% cleavage and 42.6% blastocyst development of oocytes selected for IVM. In conjunction with IVM in mTCM 199 plus 100 micrograms/ml oLH and IVC in c-SOF + NEA, efforts to eliminate protein from the fertilization medium revealed modified DM (mDM) prepared with PVA instead of BSA to be superior to TALP prepared with PVA; IVMFC data for blastocyst development were 27.4% vs. 18.2% (p < 0.05), respectively. The use of mDM for sperm preparation and IVF yielded comparable blastocyst development when either BSA or PVA was included.(ABSTRACT TRUNCATED AT 250 WORDS)
影响因子：
3.6
作者：
ROSENKRANS, CF;ZENG, GQ;FIRST, NL
通讯作者：
FIRST, NL
DOI：
10.1530/jrf.0.0910113
发表时间：
1991-01-01
期刊：
JOURNAL OF REPRODUCTION AND FERTILITY
影响因子：
--
作者：
CARNEY, EW;FOOTE, RH
通讯作者：
FOOTE, RH
影响因子：
3.6
作者：
ERBACH, GT;LAWITTS, JA;BIGGERS, JD
通讯作者：
BIGGERS, JD