Bilton R J, Moore N W
Aust J Biol Sci. 1976 Mar;29(1-2):125-9.
Goat embryos collected 5 and 7 days after mating, were cultured in vitro at 37 degrees C for 2 days, or stored at 5 degrees C for 1 or 2 days and then cultured for 2 days, or stored in liquid nitrogen (-196 degrees C) for 2-4 weeks and then cultured for 1 day. After culture some of the embryos were transferred to recipient does. Culture and storage was carried out in Dulbecco phosphate buffer enriched with 25% goat serum. 1M glycerol or 2M dimethylsulphoxide (DMSO) was added to the media used for frozen storage. Thirteen of 15 embryos cultured without prior storage showed apparently normal development in culture. Ten of the 13 were transferred and five kids were born. Twenty of 38 embryos stored at 5 degrees C developed in culture and six kids were born following the transfer of 17 embryos. Duration of storage at 5 degrees C had no marked effect upon subsequent development. Six of 48 frozen stored embryos developed in culture. All six were transferred and three kids were born.
在交配后5天和7天收集的山羊胚胎,在37℃体外培养2天,或在5℃储存1天或2天,然后培养2天,或在液氮(-196℃)中储存2-4周,然后培养1天。培养后,将一些胚胎移植到受体母羊体内。培养和储存均在添加25%山羊血清的杜尔贝科磷酸盐缓冲液中进行。用于冷冻保存的培养基中添加了1M甘油或2M二甲亚砜(DMSO)。15个未经预先储存而培养的胚胎中有13个在培养中显示出明显正常的发育。13个中的10个被移植,5只小羊出生。38个在5℃储存的胚胎中有20个在培养中发育,17个胚胎移植后有6只小羊出生。在5℃的储存时间对随后的发育没有显著影响。48个冷冻保存的胚胎中有6个在培养中发育。所有6个都被移植,3只小羊出生。