Evolution of a culture protocol for successful blastocyst development and pregnancy
Evolution of a culture protocol for successful blastocyst development and pregnancy
复制标题
DOI：
10.1093/humrep/13.1.169
复制
发表时间：
1998-01-01
影响因子：
6.1
通讯作者：
Wood, C
中科院分区：
文献类型：
--
作者：
Jones, GM;Trounson, AO;Wood, C
通过在一系列方案中测试一系列培养条件，设计了用于人胚胎发育至胚泡阶段的无细胞培养系统。所形成的培养系统具有短暂的1小时暴露于精子，然后在IVF-50培养基中培养原核合子2天。将两个或三个胚胎一起培养在20 μ l微滴的油下培养基中。然后将胚胎重新分组，并将处于相似阶段的两个或三个胚胎在油下在50 μ l Gardner’s G2培养基的微滴中一起培养3 - 5天。在第5天将胚胎转移到新鲜的G2培养基中并再培养1或2天（第6或7天）。在任何培养物中均未使用血清。胚胎移植培养基和G2培养基中添加人血清白蛋白。所有待移植给患者的囊胚的透明带均被酶促完全去除。使用该方案，52%的受精卵发育成囊胚，35名接受治疗的患者中有34名在第5天或第6天接受了82个囊胚和11个桑椹胚。在13例正在进行的妊娠（38%妊娠率/移植或37%/治疗患者）中检测到21个具有阳性心跳的胎囊（23%植入率）。我们预计，使用这种简单有效的培养系统可以进一步改善胚胎发育和选择可行的胚胎。
A cell-free culture system was designed for human embryo development to the blastocyst stage by testing a range of culture conditions in a series of protocols. The culture system that was evolved has a brief 1 h exposure to spermatozoa and then culture of the pronucleate zygote for 2 days in IVF-50 medium. Two or three embryos were cultured together in 20 mu l microdrops of medium under oil. Embryos were then regrouped and two or three at a similar stage were cultured together in 50 mu l microdrops of Gardner's G2 medium under oil from days 3 to 5. Embryos were transferred to fresh G2 medium on day 5 and cultured for a further 1 or 2 days (day 6 or 7). No serum was used in any of the cultures. The embryo transfer medium and G2 medium were supplemented with human serum albumin. The zonae of all blastocysts to be transferred to patients were completely removed enzymatically. Using this protocol, 52% of zygotes developed to blastocysts and 34 out of 35 patients treated received 82 blastocysts and 11 morulae on day 5 or 6. Twenty-one fetal sacs with positive heartbeats (23% implantation rate) were detected in 13 ongoing pregnancies (38% pregnancy rate/transfer or 37%/patient treated). We anticipate that further improvements in embryo development and the selection of viable embryos can be achieved using this simple and effective culture system.