小鼠精子微注射受精卵的体外发育与移植的研究

用显微注射方法,将精子注入小鼠卵母细胞卵周隙中,使精卵体外受精获得受精卵。并对精子注射后卵母细胞的成活率、受精率和受精卵的发育率以及移植后受胎率等方面进行了系统的研究,结果表明:卵子存活率为75.36％(1309/1737),其中264枚卵裂,受精率为20·17％(264/1309),卵裂卵经体外培养后有192枚发育到桑椹胚及胚泡期,发育率为72.73％(192/264)。将其中73枚(桑椹胚10枚,肛泡63枚)胚胎移植到10只假孕受体子宫中,一只受体妊娠产仔9只,仔鼠发育正常。     

K~ 和Ca~(2 )对牛体外受精胚胎体外发育的作用

研究观察了在 SOF PVA这一化学成分明确培养系统条件下 ,不同浓度的 K 和 Ca2 对牛体外受精胚胎体外发育的影响 ,以便为今后进一步探讨影响牛早期胚胎体外发育的因素提供实验依据 .实验 1将牛体外受精卵培养分别于含有 0、5 .0、8.3 5和 2 1 .2 m MK 的 SOF PVA培养系统中 ,结果卵裂率分别为 2 0 .0 % a、87.0 % b、80 .8% b和 85 .7% b(ae,P<0 .0 5 ) .实验 2将牛体外受精卵分别培养于含有 0、1 .1 3、1 .71和 3 .0 m MCa2 的 SOF PVA培养系统中 ,结果卵裂率分别为 0 .9% g、86.6% h、80 .9% h 和 92 .0 % h(g相似文献   

In vitro fertilization: MRC tests the climate

It is reported that Britain's Medical Research Council has published guidelines for MRC-supported research involving human embryos obtained through in vitro fertilization. In ethics committee-approved projects, researchers may use deliberately fertilized ova or, with parents' consent, fertilized ova left over from therapeutic programs. In vitro culture of human embryos is permitted only up to the implantation stage. Conditions are also set for experiments on interspecies fertilization and on freezing of embryos. Concern over physician and public reception of these guidelines has been expressed by the British Medical Association.     

Effects of different nuclear recipients on developmental potential of mouse somatic nuclear transfer embryos

Somatic cell nuclear transfer has been succeeded in procedures of nuclear transfer. One is single nucleartransfer, the other is serial nuclear transfer. Viable animals have been cloned in different species using both me-thods[1—6]. Different nuclear recipients and donors wereused in serial nuclear transfer, namely, transferring thenuclear of reconstructed embryo into enucleated MⅡoocytes[7], transferring the nuclear of reconstructed em-bryos at one cell stage into enucleated zygote[4] and t…     

体外培养时间和表皮生长因子对牦牛卵母细胞成熟及孤雌胚胎体外发育的影响

研究在成熟液中添加表皮生长因子（Epidermal growth factor,EGF）以及成熟时间对牦牛卵母细胞体外成熟及孤雌胚胎体外发育的影响,以确立牦牛卵母细胞体外成熟的最佳体系.结果表明,成熟液中添加EGF组的成熟率明显高于未添加组（P＜0.05）,并且牦牛卵母细胞的成熟率随着EGF添加的浓度增加也不断上升;随着体外培养时间的延长,成熟率逐渐增加,培养24 h后成熟率趋于稳定;胚胎体外培养液中添加EGF能显著提高孤雌胚胎的8-细胞和囊胚形成率（P＜0.05）.由此推测：在体外培养22-24 h,且添加40 μg/mL EGF最有利于牦牛卵母细胞体外成熟;胚胎培养液中添加40μg/mLEGF能显著提高牦牛孤雌胚胎的体外发育能力.     

提高供氧能力对土曲霉生产衣康酸的影响

本文研究了供氧能力对土曲霉ＩＦＯ－６３６５分批发酵生产衣康酸能力的影响。当通风比为０．５Ｖ．Ｖ．ｍ，搅拌叶轮末端线速度为１２５．７ｃｍ／ｓ时的最大衣康酸发酵产率分别是叶轮末端线速度为９４．２ｃｍ／ｓ和７８．５ｃｍ／ｓ的１．２８倍和３倍；当添加氧载体──正十二烷，叶轮末端线速度为７８．５ｃｍ／ｓ时，与对照相比，可提高产酸１４％，但在更高的叶轮末端线速度条件下，加入正十二烷，只能促进菌体的生长，对发酵产酸是抑制作用。     

兔卵输卵管培养与再移植后的成活力

本文研究分裂的兔卵在输卵管内培养与再移植。目的是观察这种胚胎的成活力,验证这种方法的实用价值。分裂期的兔卵在输卵管内培养2～3天,发育到胚泡,然后再移植。移植分自体移植和异体移植。总共再移植24例,13例母兔妊娠,产仔9窝,获仔兔21只,怀孕率54.17％,成活率25.61％.     

文昌鱼Pax1/9基因上游调控区的克隆及分析

脊椎动物Pax1/9是一重要的发育调控基因亚家族,文昌鱼基因组中仅有单一的该亚家族直系同源基因Amphi-Pax1/9,为检验此基因上游调控元件的功能在脊椎动物体内是否具有通用性,将文昌鱼Pax1/9基因上游约4.6 kb的侧翼序列与绿色荧光蛋白(GFP)报告基因连接,构建重组质粒表达载体(pAmphiPax1/9-AcGFP),显微注射斑马鱼胚胎,并以斑马鱼Pax1和Pax9的上游同源序列为阳性对照.结果表明,阳性对照斑马鱼胚胎中GFP能够表达,但注射pAm-phiPax1/9-AcGFP质粒的斑马鱼胚胎中无明显的GFP表达,这一现象可能是由于Pax1/9上游调控序列在二物种间存在较大的进化差异,启动元件具有物种特异性.     

Embryonic stem cell as nuclear donor could promote in vitro development of the heterogeneous reconstructed embryo

The nucleus of a somatic cell could be dedifferentiated and reprogrammed in an enucleated heterogeneous oocyte. Some reconstructed oocytes could develop into blastocysts in vitro, and a few could develop into term normally after transferred into foster mothers, but most of cloning embryos fail to develop to term. In order to evaluate the efficacy of embryonic stem cell as nucleus donor in interspecific animal cloning, we reconstructed enucleated rabbit oocytes with nuclei from mouse ES cells, and analyzed the developmental ability of reconstructed embryos in vitro. Two kinds of fibroblast cells were used as donor control, one derived from ear skin of an adult Kunming albino mouse, and the other derived from a mouse fetus. Three types of cells were transferred into perivitelline space under zona pellucida of rabbit oocytes respectively. The reconstructed oocytes were fused and activated by electric pulses, and cultured in vitro. The developmental rate of reconstructed oocytes derived from embryonic stem cells was 16.1%, which was significantly higher than that of both the adult mouse fibroblast cells (0%-3.1%, P < 0.05) and fetus mouse fibroblast cells (2.1%-3.7%, P < 0.05). Chromosome analysis confirmed that blastocyst cells were derived from ES donor cell. These observations show that reprogramming is easier in interspecific embryos reconstructed with ES cells than that reconstructed with somatic cells, and that ES cells have the higher ability to direct the reconstructed embryos development normally than fibroblast cells.