兔体外受精的研究

为完善家兔体外生产胚胎技术，采用FSH＋HCG法处理48只母兔，获得卵母细胞.采用获能的精子与卵母细胞进行体外受精，使用TCM199为基础液进行受精卵的体外培养.结果显示：平均每只供体兔可获得30.69枚可用卵母细胞.屠宰法获得的精子获能以后与卵母细胞进行体外受精，卵裂率达65.51%，囊胚发育率18.04%.自然射出法获得的精子获能以后，与卵母细胞进行体外受精，卵裂率达66.33%，囊胚发育率17.83%.屠宰法与自然射出法获得的精子获能以后卵裂率和囊胚发育率差异不显著.     

幼龄羊的周龄及卵母细胞体外培养体系对JIVET技术的影响

为了探讨周龄及卵母细胞体外培养体系对幼龄羊胚胎体外生产技术(JIVET)的影响,试验对10只5~10周龄杜寒杂种羔羊进行激素诱导,获取卵母细胞,分别在基础成熟液和含200μmol/L半胱氨酸的成熟液里成熟,再分别在含2%发情羊血清(ESS)的Ⅰ受精液和含3 mg/m L牛血清白蛋白(BSA)的Ⅱ受精液里受精,测定不同周龄羔羊的采卵数量、卵母细胞成熟率和卵裂率。结果表明:5,7,8,10周龄羔羊获得卵母细胞数分别为(111.00±16.97)枚/只、(139.50±28.99)枚/只、(108.50±17.68)枚/只、(42.00±11.31)枚/只,5,7,8周龄羔羊显著高于10周龄羔羊(P0.05),而5,7,8周龄羔羊之间差异不显著(P0.05);试验成熟液获得的卵母细胞成熟率比基础成熟液高3.64百分点;Ⅰ受精液获得的卵母细胞卵裂率极显著高于Ⅱ受精液(P0.01)。说明选择5~8周龄羔羊、卵母细胞体外成熟液添加一定量半胱氨酸和体外受精液含一定量ESS可以改善JIVET体系。     

不同浓度β-巯基乙醇对6周龄杜泊羊卵母细胞发育能力的影响

为了探讨成熟液里不同浓度β-巯基乙醇对6周龄杜泊羊卵母细胞发育能力的影响,试验选择6只6周龄杜泊羊,采用促性腺激素诱导活体采集卵母细胞,将获得的A、B级卵丘-卵母细胞复合体(COCs)在不同浓度(0、50、70、100μmol/L)β-巯基乙醇成熟液中成熟,再经成熟卵母细胞与获能精子共同孵育、受精卵发育培养后,统计卵裂率和囊胚率。结果表明:与对照组卵裂率(59.13%)和囊胚率(12.17%)相比,试验Ⅰ、Ⅱ、Ⅲ组的卵裂率(60.87%、63.48%、65.22%)和囊胚率(14.78%、17.39%、21.74%)均较高(P0.05),其大小顺序为试验Ⅲ组试验Ⅱ组试验Ⅰ组对照组,100μmol/Lβ-巯基乙醇组的卵裂率、囊胚率分别增加6.09个百分点和9.57个百分点。说明成熟液里添加一定量β-巯基乙醇能够提高6周龄杜泊羊卵母细胞的发育能力和受精卵质量,β-巯基乙醇浓度在100μmol/L以内,卵母细胞的卵裂率和囊胚率呈上升趋势;关于成熟液里β-巯基乙醇最佳浓度,有待进一步研究。     

荷斯坦奶牛与和牛活体采卵-体外受精（OPU/IVF）体系的建立及应用

[目的]研究牛活体采卵-体外受精（ovum pick-up and in vitro fertilization,OPU/IVF）体系,建立高效的牛体外胚胎生产系统。[方法]以从屠宰场采集的健康牛新鲜卵巢为试验材料,进行卵母细胞体外成熟、体外受精、体外胚胎培养相关条件的摸索,重点考查体外胚胎培养液中添加瘦素（leptin）对囊胚率的影响。选取13～15月龄健康荷斯坦奶牛及和牛各10头作为供体,进行活体采卵、卵母细胞体外成熟、体外胚胎生产,记录可用卵数及可用囊胚数,统计卵裂率、囊胚率;2个品种牛的体外冷冻胚胎解冻后,以荷斯坦奶牛为受体进行胚胎移植,移植后45 d统计妊娠率。[结果]添加30 U/mL的leptin可以显著（P<0.05）提高屠宰场来源牛体外胚胎的囊胚率。随机选择供体牛活体采卵,平均每头荷斯坦奶牛获得可用卵7.5枚,平均每头和牛获得可用卵8.1枚;荷斯坦奶牛及和牛的卵裂率分别为84.00%、82.71%,二者差异不显著（P>0.05）。体外培养条件下,平均每头荷斯坦奶牛获得可用囊胚4.3枚,平均每头和牛获得可用囊胚3.7枚;荷斯坦奶牛的囊胚率（57.33%）显著...     

小尾寒羊羔羊超数排卵及卵母细胞玻璃化冷冻保存效果研究

以我国地方品种小尾寒羊作为供体,对不同年龄羔羊(6～8周龄和12～14周龄)超数排卵效果以及卵母细胞冷冻保存对体外受精、体外胚胎发育、胚胎移植产羔的影响进行研究。结果表明:6～8周龄组羔羊只均超排处理获卵数和可用卵数(60.8枚和58.2枚)显著高于12～14周龄组(27.3枚和26.0枚)(P<0.05);经玻璃化冷冻—解冻后的卵母细胞体外受精,冷冻组的卵裂率和桑椹胚发育率(67.8%和35.6%)均显著降低于对照组(79.2%和53.8E)(P<0.05);玻璃化冷冻保存体外生产的胚胎,经移植后成功产下4只健康羔羊(4/52)。     

杜泊羊冷冻胚胎移植效果的影响因素及羔羊早期生长性能研究

为研究杜泊羊冷冻胚胎移植效果的影响因素以及羔羊早期的生长情况,本实验选用1 361枚质量合格的纯种黑头杜泊羊的早期冷冻胚胎作为供体,180只小尾寒羊和970只湖羊作为受体羊,采用同期发情和子宫角胚胎移植技术,研究季节(春季、秋季)、受体羊品种(小尾寒羊、湖羊)、卵巢上黄体个数(1个、2～3个)、受体羊的营养状况(中上等膘情、下等膘情)、胚胎发育阶段(桑椹胚、晚期桑椹胚、早期囊胚、囊胚和扩张囊胚)及移植胚胎数目(双胚和单胚)对于杜泊羊冷冻胚胎移植效果的影响。结果显示:秋季(11月)同期发情率显著高于春季(4月),但妊娠率没有显著差异;在春季,不同品种受体羊的同期发情率和妊娠率无显著性差异;受体羊卵巢上黄体个数对妊娠率无显著影响;中上等膘情受体羊的妊娠率显著高于下等膘情受体羊;移植扩张囊胚的受体羊妊娠率为30.0%,显著低于其他4个胚胎发育阶段;移植双胚的受体羊妊娠率低于移植单胚的受体羊。羔羊早期生长发育指标统计结果表明:在潍坊地区,小尾寒羊和湖羊均可以作为胚胎移植的受体,但是小尾寒羊体型偏大,每日饲料消耗偏多,湖羊作为受体羊在节约饲料方面更有优势。     

半胱胺对羔羊卵母细胞体外成熟及胚胎发育的影响

实验旨在研究半胱胺对4～8周龄哈萨克羔羊卵母细胞体外成熟和发育能力的影响。在体外成熟培养基中添加不同浓度的半胱胺(0、50、100、200μmol/L),统计卵母细胞卵裂率、囊胚率、成熟率、正常受精率及卵裂和胚泡率,并用SPSS 17.0进行方差分析。结果表明：与空白组相比,添加100μmol/L半胱胺可提高羔羊卵母细胞卵裂率、囊胚率、成熟率、正常受精率、卵裂率及胚泡率(P<0.05),添加100μmol/L半胱胺可提高卵母细胞的正常受精率和滋养层细胞数(P<0.05)。可见,100μmol/L半胱胺可显著改善羔羊卵母细胞的体外成熟、受精和发育能力。     

卵巢质量和保存液对牛卵母细胞体外发育能力的研究

为研究卵巢质量和保存液对牛卵母细胞孤雌发育能力的影响,本实验将采集卵巢按其质量分为A、B 2组,于生理盐水中运输到实验室,利用切割法采集卵巢表面卵泡中的卵母细胞培养,成熟后孤雌激活;将采集牛卵巢随机分到D-PBS保存液与生理盐水中,运送到实验室后采集卵母细胞培养。以卵母细胞的成熟率、孤雌胚的卵裂率、囊胚率来评价卵巢质量、保存液对卵母细胞发育能力的影响。结果表明:A组卵巢平均获得的卵母细胞数(2.1枚)显著低于B组卵巢获得的卵母细胞数(5.5枚),从A组卵巢上获得的卵母细胞成熟率(56.98%)、卵裂率(42.86%)及囊胚率(9.52%)显著低于B组卵巢上获得的卵母细胞成熟率(84.38%)、卵裂率(70.17%)及囊胚率(31.50%)(P<0.05);使用含离子丰富的D-PBS溶液,较对照组保存液生理盐水,其卵母细胞的成熟率、卵裂率及囊胚率均没有显著差异(P>0.05)     

封闭式拉长细管冷冻牛卵母细胞的研究

为探索封闭式拉长细管(closed pulled straw,CPS)冷冻牛卵母细胞的效果,将牛卵母细胞分别采用细管、开放式拉长细管(open pulledstraw,OPS)和CPS 3种方法进行冷冻,解冻后进行体外成熟、体外受精和胚胎体外培养。采用CPS法分别对GV期和成熟(MII期)牛卵母细胞进行冷冻保存,解冻后将卵母细胞培养至成熟,进行体外受精和胚胎体外培养。结果显示:OPS组和CPS组卵母细胞的正常形态率分别为83.1%和77.8%,成熟率分别为66.9%和64.4%,卵裂率分别为45.8%和43.4%,囊胚率分别为6.6%和6.0%,组间上述指标差异均不显著(P>0.05);但OPS和CPS组上述4项指标均显著高于细管冷冻组(P<0.05)。解冻后GV期和MII期卵母细胞卵裂率分别为44.5%和57.3%,囊胚率分别为6.8%和17.9%,组间卵裂率和囊胚率差异均显著(P<0.05)。结果表明:CPS法既具有OPS法快速降温的优点,同时,还能避免卵母细胞和液氮直接接触,降低卵母细胞通过液氮被污染的风险。因此,CPS冷冻法可以用于牛卵母细胞的冷冻。     

换液培养对猪卵母细胞的成熟及胚胎体外发育的影响

通过后期去除激素培养比较了卵丘卵母细胞成熟及构建克隆胚后的发育情况,同时孤雌胚胎和体外受精胚胎体外培养48 h后全量换液,比较了其卵裂率和囊胚率之间的差异。结果显示,卵丘卵母细胞培养22h后换成不含激素的成熟培养液继续培养至44 h,其成熟率与对照组无显著差异(P0.05);后期克隆胚胎卵裂率和囊胚率与对照组均无显著差异(P0.05)。孤雌胚胎换液培养卵裂率高于未换液组,囊胚率低于未换液组,但差异均不显著(P0.05)。体外受精胚胎换液培养卵裂率以及囊胚率和未换液组差异不显著(P0.05)。本实验结果说明换液培养对卵母细胞的成熟及后续的体细胞克隆胚发育无显著影响,孤雌和体外受精胚胎换液培养对后期发育也无显著影响。     

Cryopreservation of immature oocytes of the indigeneous Vietnamese Ban Pig

We report the cryopreservation of oocytes from Ban miniature pigs which are endemic in Vietnam. Immature cumulus‐oocyte complexes were collected from antral follicles of 7–8 mo old female cyclic Ban pigs and vitrified in micro‐drops. Oocyte morphology, lipid content, post‐warming survival, nuclear maturation, and embryo development were compared to those of oocytes from commercially slaughtered Landrace × Large white hybrid pigs. The size of oocytes in the two breeds was similar. However, significantly lower amounts of intracellular lipid were detected in Ban oocytes. There was no difference (p > 0.05) between Ban and Landrace × Large white oocytes in percentages of post‐warming survival (93.1 ± 3.4% vs. 70.7 ± 16.7%, respectively) and nuclear maturation after in vitro maturation (80.4 ± 5.1% vs. 90.0 ± 1.3% respectively). Similarly, cleavage (30.8 ± 7.8% vs. 10.3 ± 6.1%, respectively) and blastocyst development rates (9.4 ± 5.0% vs. 0.79 ± 0.79, respectively) were not different (p > 0.05) between vitrified Ban and Landrace × Large white oocytes after in vitro fertilization and embryo culture. In conclusion, high survival and maturation rates were achieved after vitrification of immature Ban oocytes and their cryo‐tolerance was similar to that of Landrace × Large white oocytes, despite the difference in lipid content. We succeeded to generate reasonable rates of blastocysts from vitrified Ban oocytes by in vitro fertilization.     

Effect of Antioxidants During Bovine In Vitro Fertilization Procedures on Spermatozoa and Embryo Development

Increased amounts of reactive oxygen species (ROS) during in vitro fertilization (IVF) may cause cytotoxic damage to gametes, whereas small amounts of ROS favour sperm capacitation. The aim of this study was to investigate the effect of antioxidants [50 μ m β-mercaptoethanol (β-ME) and 50 μ m cysteamine (Cyst)] or a pro-oxidant (5 m m buthionine sulfoximine) on the quality and penetrability of spermatozoa into bovine oocytes and on the subsequent embryo development and quality when added during IVF. Sperm quality, evaluated by the integrity of plasma and acrosomal membranes, and mitochondrial function, was diminished (p < 0.05) after 4-h culture in the presence of antioxidants. Oocyte penetration rates were similar between treatments (p > 0.05), but antioxidants adversely affected the normal pronuclear formation rates (p < 0.05). The incidence of polyspermy was high for β-ME (p < 0.05). No differences were observed in cleavage rates between treatments (p > 0.05). However, the developmental rate to the blastocyst stage was adversely affected by Cyst treatment (p < 0.05). The quality of embryos that reached the blastocyst stage, evaluated by total, inner cell mass (ICM) and trophectoderm cell numbers and ICM/total cell ratio was unaffected (p > 0.05) by treatments. The results indicate that ROS play a role in the fertilizing capacity in bovine spermatozoa, as well as in the interaction between the spermatozoa and the oocytes. It can be concluded that supplementation with antioxidants during IVF procedures impairs sperm quality, normal pronuclear formation and embryo development to the blastocyst stage.     

Production of Lambs After the Transfer of Fresh and Cryopreserved in vitro Produced Embryos from Prepubertal Lamb Oocytes and Unsorted and Sex-sorted Frozen-thawed Spermatozoa

Cumulus-oocyte complexes from hormone-stimulated 3-4-week-old (n=43) and 6-7-week-old (n=12) prepubertal lambs were matured in vitro and incubated with unsorted, or X- or Y-spermatozoa separated with a high-speed cell sorter (SX MoFlo)frozen-thawed. Presumptive zygotes were then cultured to the blastocyst stage, and transferred to recipients fresh or after cryopreservation (frozen). Oocyte cleavage was higher (p <0.05) with unsorted (515/926, 55.6%) than X- or Y-spermatozoa (261/672, 38.8% and 229/651, 35.2%, respectively) and blastocyst formation (% zygotes) by Day 9 of in vitro culture was lower (p <0.05) for X- (102/261, 39.1%) than unsorted spermatozoa (249/515, 48.3%), but did not differ between Y-spermatozoa (103/229, 45.0%) and unsorted spermatozoa, or between X- and Y-spermatozoa (p >0.05). For fresh embryos, survival to term was 50.0% (3/6) for unsorted, 0.0% (0/6) for X- and 16.7% (1/6) for Y-spermatozoa-derived embryos (p >0.05), and for frozen embryos was 4.0% (2/50) for unsorted, 9.1% (2/22) for X- and 2.9% (1/34) Y-spermatozoa-derived embryos (p >0.05). Of the two lambs born from X-spermatozoa-derived embryos, one was female (50%), and from the two Y-spermatozoa-derived lambs, both were male (100%), demonstrating that lambs can be produced after the transfer of fresh and cryopreserved IVP embryos derived from prepubertal lamb oocytes and frozen-thawed sex-sorted sperm.     

Effect of Inter‐Breed Embryo Transfer on Lamb Growing Performance and Survival

The roles of recipient and embryo genotype in determining the growth performance and survivability of offspring based on specific behavioural differences were investigated using inter‐breed embryo transfer. This study was carried out using three recipient genotypes (Awassi, Redkaraman and Tuj) and two embryo genotypes (Charollais and Romanov) to obtain the six possible combinations of ewe and lamb genotypes. Data were collected from 71 recipient ewes (10 Redkaraman with Charollais and 15 Redkaraman with Romanov embryos; 10 Tuj with Charollais and 12 Tuj with Romanov embryos, and 12 Awassi with Charollais lambs and 12 Awassi with Romanov embryos); all ewes received two frozen‐thawed embryos. Awassi ewes had a significantly longer duration of the licking/grooming event (25.5 min, p < 0.05) than Tuj ewes. Charollais lambs were significantly (p < 0.05) more likely to require birth assistance compared to Romanov lambs. Romanov lambs were significantly more (p < 0.01) active than Charollais lambs in the first 2 h after birth; ewe breed had no effect on lamb behaviour. There was no recipient breed effect on either birth or weaning weights of lambs. Charollais lambs were recorded with higher birth (5.5 ± 0.3 kg vs 3.9 ± 0.2 kg; p < 0.001) and weaning (29.4 ± 1.2 kg vs 22.4 ± 1.9 kg; p < 0.001) weights compared to Romanov lambs. At weaning Romanov lambs had significantly higher (95% vs 75%; p < 0.05) survival rates, however, this was not significantly affected by recipient breed. It was concluded that recipient breed was not an important factor in survival and weaning performance of embryo transferred lambs from a prolific breed (Romanov) while these traits recorded for lambs from meat type (Charollais) embryos were influenced by dam breed.     

湖羊胚胎和精液超低温保存效果的评价

【目的】 检验长期保存在国家家畜基因库的湖羊冷冻胚胎和冷冻精液的质量,评价超低温冷冻保存技术保种的效果。【方法】 对保存于国家家畜基因库的冷冻胚胎（保存20年）和冷冻精液（保存20和30年）进行复苏,进行相应鉴定后分别进行胚胎移植和人工授精,同时以0年冷冻精液和新鲜精液为对照,测定其受胎率和后代的羔皮性能、生长性能及繁殖性能,检验保存效果。【结果】 本试验共解冻胚胎36枚,其中,A级胚胎22枚,B级胚胎7枚,C级胚胎5枚,D级胚胎2枚,冷冻胚胎复苏利用率达到94.44%（34/36）,A级胚胎率达到61.11%（22/36）,移植34枚,产羔17只,冷冻胚胎复苏移植产羔率达50.00%;保存30年的冷冻精液复苏后平均精子活力达35%,受胎率为58.57%,平均产羔1.90只;保存20年冷冻精液复苏后平均精子活力达31%,受胎率为53.66%,平均产羔1.95只;胚胎复苏移植羊、30和20年冷冻精液后代体型外貌鉴定均符合纯种湖羊特征,没有畸形个体;羔羊一级羔皮率分别是31.25%、42.03%和23.81%,保持了当年湖羊的羔皮特性;生长发育及繁殖性能与现有湖羊群体性能基本保持一致。【结论】 利用超低温冷冻技术长期保存湖羊冷冻胚胎和冷冻精液的方法是可行的,对地方品种保种具有重要意义。     

杜泊、无角陶赛特、萨福克羊与湖羊杂交F1代羔羊育肥性能研究

为进一步研究不同肉羊品种与湖羊杂交F₁代羔羊育肥效果,本试验选择国外优良的肉羊品种杜泊羊、无角陶赛特羊和萨福克羊作父本,国内优良地方品种湖羊作母本开展杂交试验研究,所获得的杂交F₁代羔羊在全舍饲条件下进行育肥。测定杂交F₁代羔羊初生重、不同月龄体重和平均日增重等生长性能。结果表明：试验A组（杜泊公羊与湖羊母羊杂交F₁羔羊）、试验B组（无角陶赛特公羊与湖羊母羊杂交F₁羔羊）、试验C组（萨福克公羊与湖羊母羊杂交F₁羔羊）6月龄平均重分别为44.76±3.62 g、 43.65±4.18 g、43.28±3.34 g,均极显著高于对照组D 6月龄平均重36.32±3.86 g,差异极显著(P＜0.01)。试验A组、B组、C组与对照组D相比较,试验A组、B组、C组每只羊的利润较对照组分别多收入80.02元、69.74、42.74元,差异极显著（P＜0.01）,其中试验A组的生长性能和利润均优于试验B组和C组,差异显著(P＜0.05)。     

Maturation Status, Protein Synthesis and Developmental Competence of Oocytes Derived from Lambs and Ewes

The efficacy of oocyte selection for in vitro embryo production depends on the abundance and diameter of follicles, cumulus layers around the oocytes and subsequent fertilization. Application of `ovum pick-up' technique allows us to utilize partially matured oocytes for embryo production even from juvenile subjects. To compare their developmental competence, oocytes derived from lambs and ewes and cultured in maturation medium for up to 26 h were assessed at 2 h intervals by confocal microscopy after chromatin and microtubulin-specific fluorochrome labelling. Lamb oocytes reached second meiotic metaphase (MII) at lower numbers at 24 h (60.0%) and 26 h (28.6%) whereas 85.7% of adult-derived oocytes attained MII status by 24 h of maturation. Radiolabelling of oocyte proteins revealed higher incorporation of [³⁵S-]-methionine and [³⁵S]-cysteine in adult-derived oocytes compared to lamb oocytes. Although the cleavage rate of lamb oocytes was similar to that of ewe oocytes, the proportion reaching blastocyst stage was significantly lower (p < 0.05) in the lamb-derived oocytes. However, blastocysts from both types of oocytes displayed similar cell lineage allocations to inner cell mass and trophectoderm.     

不同品种牛卵泡液对水牛卵母细胞体外受精效果的影响

试验利用水牛卵泡液(BuFF)和黄牛卵泡液(BoFF)对不同来源水牛卵母细胞体外受精效果的影响进行了探讨,以完善水牛体外受精培养系统,进一步提高水牛胚胎体外生产效率。试验按成熟培养液中添加卵泡液替代胎牛血清量共分4个组。不添加卵泡液(0%+10%胎牛血清)为Ⅰ组(对照组);添加5%卵泡液+5%胎牛血清为Ⅱ组;添加10%卵泡液+0%胎牛血清为Ⅲ组;添加15%卵泡液+0%胎牛血清为Ⅳ组。结果表明,添加BuFF对活体采集卵母细胞和屠宰场收集卵母细胞的体外受精卵分裂率无显著影响(P0.05),但添加5%和10%BuFF对卵母细胞体外受精后的胚胎发育有明显促进作用,囊胚率均极显著高于对照组和15%BuFF组(P0.01),5%和10%BuFF组间无显著差异(P0.05);添加15%BuFF囊胚率有降低的趋势,但与对照组相比差异不显著(P0.05)。而添加10%BoFF组活体采集卵母细胞体外受精的受精卵分裂率和囊胚率均极显著高于对照组和5%BoFF组(P0.01),添加5%BoFF组的受精卵分裂率和囊胚率与对照组无显著差异(P0.05);添加BoFF对屠宰场收集水牛卵母细胞体外受精卵分裂率无显著差异(P0.05),但添加5%和10%BoFF组的囊胚率均显著高于对照组和15%组(P0.05),添加15%BoFF组与对照组相比,囊胚率显著降低(P0.05),5%和10%BoFF组间囊胚率无显著差异(P0.05)。综合以上结果,在水牛卵母细胞成熟培养液中添加5%～10%的BuFF或BoFF代替牛血清,可明显提高水牛体外胚胎生产效率,且以添加同种的BuFF效果略好。     

不同月龄绵羔羊卵泡发育的比较研究

本研究旨在比较1、3月龄绵羔羊激素处理后卵巢、子宫及血液中促卵泡激素(FSH)变化,研究不同月龄对羔羊卵泡发育的影响。通过对1、3月龄羔羊进行FSH和孕马血清促性腺激素(PMSG)处理,比较羔羊在激素处理前后血液中FSH水平和卵巢、子宫大小变化,卵巢上2～8 mm卵泡的数量和卵母细胞体外成熟、受精后胚胎的发育情况。结果表明:1月龄羔羊实验组体内整体FSH水平高于3月龄羔羊(P<0.05)。1月龄羔羊注射外源激素后两侧卵巢可获卵母细胞数(42.3±2.5、36.8±1.1)枚及体外受精囊胚发育率(16.33%±0.96%)显著高于3月龄羔羊卵母细胞数(10.0±0.7、8.5±0.6)枚及囊胚率(9.29%±1.55%)(P<0.05)。羔羊进行超数排卵处理时卵巢上卵泡发育与血液中FSH水平密切相关,且较高的FSH水平预示着较多的卵泡发育。     

Effect of Oocytes in vivo and in vitro on the Developmental Potential of Porcine Somatic Cell Cloned Embryos

To investigate the impact of porcine oocytes in vivo and in vitro maturation (IVM) on the development of porcine somatic cell cloned embryos,the somatic cell cloned embryos cultured in vitro and the sows were treated with hormones to collect mature oocytes in vivo,and the cleavage rate, blastocyst rate and embryo implantation were compared. The results showed that the average number of ovulation in PGC+PMSG+HCG group was significantly higher than that of PGC+HCG,PMSG+HCG and the natural estrus groups (P<0.05). The oocytes collected in vivo could be used for the construction, and the available oocytes rate reached more than 90%,and there was no significant difference among the four groups (P>0.05),which indicated that groups treated by hormone could obtain more available oocytes and the quality of oocytes was not significant different. In vivo and in vitro matured oocytes were used as nuclear transfer embryos of recombinant receptor,the fusion efficiency (80.31% and 79.29%) and cleavage rate (90.40% and 86.51%) were not significant different (P>0.05), but the proportion of in vivo matured oocytes cloned embryos developed into the blastocyst stage was significantly higher (P<0.05). The reconstructed embryos made from in vivo and in vitro matured oocytes were transplanted into surrogate sows (transferred 30 or 60 embryos),10 piglets were born in in vivo maturation of cloned embryo transfer group,while there was no implantation in in vitro maturation of cloned embryo transfer group. The results showed that high quality oocytes obtained by superovulation could significantly increase the blastocyst rate of embryos,reduce the number of embryos transferred and improve the pregnancy rate of surrogate sows.