6—DMAP对小鼠卵母细胞减数分裂启动及孤雌发育作用

小鼠卵泡卵母细胞体外培养过程中加入２ｍｍｏｌ／Ｌ６－ＤＭＡＰ可抑制卵母细胞自发的染色持浓缩和生发泡破裂（ＧＶＢＤ）。源自超排的ＭⅡ期卵母细胞则能为６－ＤＭＡＰ所激活。ｈＣＧ注射后１８－１９ｈ的卵母细胞置于２ｍｍｏｌ／Ｌ６－ＤＭＡＰ的ＣＺＢ溶液中培养０．５ｈ、１ｈ、２ｈ、３ｈ,卵母细胞的激活率分别为２６．１％、７５．２％、７５．８％、７７．３％、卵裂率分别为８８．２％、７３．２％、６７．０％、５８．     

促性腺激素对猪卵丘细胞-卵母细胞复合体体外减数分裂恢复作用的研究

本实验利用猪卵母细胞体外无血清培养技术,选用猪卵泡液中自然存在的次黄嘌呤（ＨＸ）作为卵母细胞自发成熟的抑制剂,研究了促性腺激素对猪卵丘细胞－卵母细胞复合体（ＣＥＯ）减数分裂恢复的具体作用。ＣＥＯ在含有不同浓度的促性腺激素（ＦＳＨ,ｈＣＧ,ＦＳＨ＋ｈＣＧ）的培养液中培养２４ｈ,观察卵母细胞减数分裂恢复（ＧＶＢＤ）情况。实验结果如下：１．ＦＳＨ（１－５００ＩＵ／Ｌ）能够明显刺激ＣＥＯ克服ＨＸ的抑制作用而恢复减数分裂（Ｐ＜０．０５）,该作用具有剂量依赖性;２．ｈＣＧ（１－５００ＩＵ／Ｌ）对ＣＥＯ减数分裂的恢复无明显作用;３．ｈＣＧ（１０－５００ＩＵ／Ｌ）与ＦＳＨ（１０,１００ＩＵ／Ｌ）无协同作用。上述结果表明,猪ＣＥＯ减数分裂的恢复可能主要依赖于ＦＳＨ的作用,该作用能使猪卵丘细胞产生一种或几种阳性因子,作用于卵母细胞,从而克服ＨＸ的抑制作用而恢复减数分裂。ｈＣＧ无明显作用,可能是因为卵丘细胞上没有ＬＨ受体或ＬＨ受体的数量不足     

促性腺激素对猪卵丘细胞—卵母细胞复合体体外减数分裂？…

本实验利用猪卵母细胞体外无血清培养技术,选用猪卵泡液中自然存在的次黄嘌呤（ＨＸ）作为卵母细胞自发成熟的抑制剂,研究附了中性腺激素对猪卵丘细胞－卵母细胞复体（Ｃ ＥＯ）减数分裂恢复的具体作用。ＣＥＯ在含有不同浓度的促性腺激素（ＦＳＨ,ｈＣＧ,ＦＳＨ＋ｈＣＧ）的培养液中培养２４ｈ,观察卵母细胞减数分裂恢复（ＧＶＢＤ）情况。实验结果如下：１．ＦＳＨ（１－５００ＩＵ／Ｌ）能够明显刺激ＣＥＯ克服ＨＸ的抑制作     

水稻叶片中过氧化氢与核酮糖—1,5—二磷酸羧化酶／加氧酶降解 …

甲基紫精（ＭＶ）处理水稻植株能快速诱导核酮糖－１,５－二磷酸羧化酶／加氧酶（Ｒｒｂｉｓｃｏ,ＥＣ４．１．１．３９）及其它可溶性蛋白的降解。ＭＶ浓度越高,降解速率越高。ＭＶ能诱发叶片内源Ｈ２Ｏ２迅速积累。光合电子传递抑制剂ＤＣＭＵ（１５０μｍｏｌ／Ｌ）能显著抑制ＭＶ（１００μｍｏｌ／Ｌ）诱导的Ｒｕｂｉｓｃｏ及其它可溶性蛋白的降解;活性氧清除剂抗坏血酸（５ｍｍｏｌ／Ｌ）、甘露醇（１０ｍｍｏｌ／Ｌ）、苯     

铝和铝＋钙对小麦幼苗根尖质膜,液泡膜微囊ATP酶和膜流动性？…

研究了铝和铝＋＿钙对小麦功苗根尖质膜、液泡膜微囊Ｈ＾＋－ＡＴＰ酶、Ｃａ＾２＋－ＡＴＰ酶、Ｍｇ＾２＋－ＡＴＰ酶活性及共动力学参数和膜流动性的影响。在质膜和液泡膜微囊制剂中加入１．０ｍｍｏｌ／Ｌ的ＡＩ＾３＋（ＡＩＣＩ３）时,Ｈ＾２＋－ＡＴＰ囊制剂中加入１．０ｍｍｏｌ／Ｌ的ＡＩ＾３＋（ＡＩＣＩ３）时,Ｈ＾＋－ＡＴＰ酶、Ｃａ＾２＋－ＡＴＰ酶、Ｍｇ＾２＋－ＡＴＰ酶活笥和酶促反应的Ｖｍａｘ及膜流动性下降,而酶     

兔血管平滑肌延迟整流钾通道研究及其与克隆Kv1.5通道的电生理特性比较

本文用膜片箝全细胞技术比较了研究了单个兔肺动脉血管平滑肌细胞上延迟整流钾通道与克隆Ｋｖ１．５通道的电生理及药理学特性。将平滑肌细胞箝制在－４０ｍＶ,以１０ｍＶ的步跨阶跃去极化（０￣６０ｍＶ）可产生一系列快速上升的外向电流,几无衰减,其激活曲线的Ｖ１／２为２７．２ｍＶ。灌流液中加入１００ｍｍｏｌ／Ｌ和ＴＥＡ １ｍｍｏｌ／Ｌ ４ＡＰ,电流幅度均明显减小,细胞外Ｃａ＾２＋水平由１．５ｍｍｏｌ／Ｌ降至０．     

锂对小鼠高增殖潜能集落形成细胞和粒巨噬系祖细胞体外增殖的影响

本文观察了锂对ＢＡＬＢ／Ｃ小鼠骨髓高增殖潜能集落形成细胞（ＨＰＰ－ＣＦＣ）和粒巨噬系祖细胞ＣＦＵ－ＧＭ体外增殖的影响。ＨＰＰ－ＣＦＣ集落由ＩＬ－１、ＩＬ－６、ＷＥＨＩ３条件培养液（ＷＥＨＩ３－ＣＭ,含有ＩＬ－３）及Ｌ９２９条件培养液（Ｌ９２９－ＣＭ,含有Ｍ－ＣＳＦ）所支持,而ＣＦＵ－ＧＭ由ＷＥＨＩ３－ＣＭ所支持。结果显示,ＬｉＣｌ浓度在０．４－２ｍｍｏｌ／Ｌ时呈现剂量依赖性抑制ＨＰＰ－ＣＦＣ增殖;而在０．４－１ｍｍｏｌ／Ｌ的浓度范围内,则对ＣＦＵ－ＧＭ的增殖起剂量依赖性促进作用。这些结果提示ＬｉＣｌ对ＨＰＰ－ＣＦＣ和ＣＦＵ－ＧＭ的作用不同,可能锂有诱导ＨＰＰ－ＣＦＣ向成熟细胞分化的作用     

细胞外Mn^2＋对内向整流钾通道（IRK1）的阻断作用

目的和方法：采用双微电极电压钳（ＴＥＶ）法研究细胞外Ｍｎ＾２＋对非洲爪蟾卵母细胞表达的内向整流钾通道（ＩＲＫ１）的阻断作用。结果：细胞外Ｍｎ＾２＋浓度分别为１、１．２５、２．５、５、１０和２０ｍｍｏｌ／Ｌ,Ｋ＾＋浓度为９０ｍｍｏｌ／Ｌ,可见Ｍｎ＾２＋对ＩＲＫ１的瞬间电流（旋加电压后２ｍｓ）具有Ｍｎ＾２＋浓度依赖性和电压依赖性阻断作用;细胞外加Ｍｎ＾２＋浓度较高时强;细胞外Ｋ＾＋浓度为９０ｍｍｏｌ／     

氧代赖氨酸抗真菌作用机制的初步研究

氧代赖氨酸对致病真菌类秃发念珠菌的ＭＩＣ为０．８－３．１μｇ／ｍｌ,而两性霉素Ｂ为１．６－３．１μｇ／ｍｌ．氧代赖氨酸高于赖氨酸１０倍浓度,无明显抑制白含珠菌对Ｌｙｓｉｎｅ的吸收。Ｏｘａｌｙｓｉｎｅ０．４ｍｍｏｌ／Ｌ对白色念珠菌细胞膜透性,如胞内物于ＵＶ２６０ｎｍ下的吸收物质外泄无明显影响。放射性前体物渗入试验结果指出,Ｏｘａｌｙｓｉｎｅ０．４ｍｍｏｌ／Ｌ弱抑制（＾１４Ｃ）－Ｍｅｔｈｉｏｎｉｎ和（     

紫细菌RS601DCPIPH2→MV电子传递的研究

紫色非硫细菌菌株Ｒｂ．ｓｐｈｅｒｏｉｄｅｓ６０１（ＲＳ６０１）的载色体具有典型的Ｒｂ．ｓｐｈａｅｒｏｉｄｅｓ光谱特性。在光照和以ＤＣＰＩＰＨ２为电子供体条件下,ＲＳ６０１载色体能够串联呼吸电子传递链而耗氧或还原ＭＶ而构成ＤＣＰＩＰＨ２→ＭＶ的非循环电子传递链。电子传递抑制剂ＯＰ抑制ＤＣＰＩＰＨ２→ＭＶ电子传递活性,其中Ｉ５０为１．０ｍｍｏｌ／Ｌ;抗霉素Ａ对该电子递影响较小,不存在Ｉ５０。ＬＤＨＸ,     

绵羊卵母细胞体外核成熟抑制及其对胚胎发育潜力的影响

本研究旨在探讨次黄嘌呤 +dbcAMP或IBMX +dbcAMP对绵羊卵母细胞体外成熟的可逆性抑制作用 ,以及这种抑制作用对卵母细胞胚胎发育潜力的影响。自绵羊卵巢分离卵丘 -卵母细胞复合物进行体外培养 ,培养基中分别加入或不加入上述抑制剂。培养 6h后各组取部分卵母细胞固定染色检查卵母细胞核成熟情况 ;将其余的卵母细胞分别移入无抑制剂的成熟培养基中继续培养 18h后 ,再次检查各组卵母细胞核成熟情况 ,并进行体外受精和胚胎培养。结果表明 :次黄嘌呤 +dbcAMP或IBMX +dbcAM都分别能使 6 0 %以上的绵羊卵母细胞抑制在GV期。这种抑制是可逆性的 ,去除抑制剂后卵母细胞能恢复减数分裂 ,并加快由GVBD到MⅡ的成熟过程。各处理组受精率、卵裂率和囊胚发育率与对照组相比无显著性差异 ,表明卵母细胞的胚胎发育潜力没有受损。上述物质对卵母细胞成熟的可逆性抑制可用于研究卵母细胞成熟及其胚胎发育潜力的调节机制。     

Factors affecting electrical activation of porcine oocyte matured in vitro

This work was undertaken to improve conditions for in vitro maturation and activation of porcine oocytes. Experiments were designed to compare: (i) electrical pulse frequency, (ii) methods of oocyte preparation, (iii) maturation conditions, and (iv) electrical poration medium on development. Oocytes were harvested by follicle dissection or aspiration, co-cultured with follicle shells in M199 based medium with or without media changes at 38.5°C in 5% CO₂ under non-static conditions for 48 h and electroactivated using single or multiple pulses (current strength 1.0 kV/cm for 50 μs in 0.28 M inositol or mannitol based media with 10 mM histidine) at different time intervals. The results showed: (i) neither the pulse frequency nor the pulse interval influenced rates of pronuclear formation but multiple pulse activation (3 pulses at 5 min intervals) induced a higher incidence of development and progression through the 4-cell block in contrast to one pulse activation; (ii) both the rate of nuclear maturation (88.6% vs. 77.6%) and post-activation cleavage (89.8% vs. 67.4%) were higher (P < 0.05) when oocytes were collected by follicle dissection rather than by aspiration; (iii) while changing to a hormone-free medium at 24 h was without effect on maturation (91.9% vs. 91.7%), rate of cleavage (81.6% vs. 72.3%, P < 0.05) at 24 h was enhanced by the medium change; and (iv) oocytes activated with 3 pulses 5 min apart in mannitol based medium at 48–49 h and at 53–54 h formed pronuclei at a comparable rate but subsequent parthenogenetic development was higher in the older eggs. By contrast, inositol-based medium supported development of young and old eggs equally well. Calcium and magnesium ions are, however, necessary in both mannitol and inositol media for activation of porcine oocytes matured in vitro. The present results suggest that optimal parthenogenetic activation and early development of IVM pig oocytes could be obtained if oocytes are harvested by dissection, cultured for 24 h in hormone-containing medium before being placed in hormone free medium and activated at 48 h in inositol based medium using a three pulse activation system.     

不同活化方法对小鼠卵母细胞孤雌发育的影响（简报）

In order to study effects of electro-fusion and strontium chloride (SrCl2) activation in nuclear transfer experiment on activation and development of mouse oocytes, concentration and treatment duration of SrCl2, electro-pulse and electro-pulse combining SrCl2 were used to activate mouse oocytes which were obtained after hCG 17h. Activated oocytes were cultured in vitro in CZB medium. The results were as follows: 82.4% activation percentage was obtained when the oocytes were treated with 10mmol/L SrCl2 for 6h, it was significantly (P>0.05) higher than those obtained from that treated with the 5mmol/L or 10mmol/L SrCl2 for 4h. The activation percentage was not significantly different between 5mmol/L and 10mmol/L SrCl2 for 6h, but the percentage of morula and blastocyst in 10mmol/L SrCl2 6h group was significantly (P > 0.05) higher than those in 5mmol/L SrCl2 6h group. In the groups of treatment with electro-pulse, the best activation percentage (70.9%) was obtained when the oocytes were treated with 1.0kv/cm, 320micros, 3 pulses, but M + B percentage (7.9%) was low. In the groups of treatment with electro-pulse combining with SrCl2, the best result was acquired (activation and M + B percentage were 75.0% and 57.3% separately) when the oocytes were treated in 10mmol/L SrCl2 for 6h interval 1h after treated with 1.8kv/cm, 10s, 1pulse. These results show that the treatment with electro-pulse combining SrCl2 is a better way to mouse parthenogenesis.     

体外培养条件下促性腺激素对昆明白小鼠卵母细胞成熟及卵丘扩展的影响

研究促卵泡激素（FSH）,人绒毛膜促性腺激素（hCG)对昆明白小鼠卵母细胞成熟和卵丘扩展的影响,以及体外培养时卵丘扩展与卵母细胞成熟之间的关系,FSH可以明显促进次黄嘌吟（HX）抑制条件下的卵丘－卵母细胞复合体CEO卵母细胞成熟及卵丘扩展,其最佳作用剂量为100IU／L,且FSH作用30分钟即可以使CEO获得恢复减数分裂的信息,在HX存在的条件下,FSH处理后10hr,CEO卵丘明显扩展,而生发泡破裂（GVBD）则在16－20hr明显增加,所有卵丘未扩展的CEO中卵母细胞均未发生GVBD,低剂量hCG单独或与FSH共同存在,对CEO卵母细胞成熟及卵丘扩展均无明显影响;高剂量hCG可以部分抑制FSH对卵母细胞成熟的促进作用,结果表明：FSH明显促进CEO卵母细胞成熟及卵丘扩展,而hCG却不具有此作用,体外培养条件下（含次黄嘌呤）,卵丘扩展是卵母细胞成熟的前提条件,但卵母细胞成熟并不需要卵丘完全扩展。     

应用氯化锶和放线菌酮对小鼠卵母细胞进行孤雌活化的研究

本试验研究了SrCl_2浓度和作用时间,以及卵龄和蛋白合成抑制剂放线菌酮等对昆明种小鼠卵母细胞活化的影响。研究表明,以含1.6mmol/L SrCl_2的无钙M16液对小鼠卵母细胞活化效果最好(87.0％),显著(P<0.05)优于SrCl_2浓度为1.0、5.0、10.0mmol/L的同种液体。SrCl_2作用时间10分钟显著(P<0.05)好于5、20、30或60分钟。注射hCG后18和20小时卵母细胞的活化率(分别为87.0％和84.6％)显著(P<0.01)高于14或16小时的活化率(分别为4.8％和16.5％)。CHX与SrCl_2联合使用产生显著的协同促进卵母细胞活化作用。     

Conditions for in vitro maturation and artificial activation of ferret oocytes

The ferret represents an attractive species for animal modeling of lung diseases because of the similarity between ferret and human lung biology and its relatively small size and short gestation time. In an effort to establish experimental protocols necessary for cloning ferrets, optimized conditions for in vitro maturation and artificial activation of ferret oocytes were examined. Cumulus-oocyte complexes were harvested from ovaries of superovulated ferrets, and in vitro maturation was evaluated in three different culture media: medium 1 (TCM-199 + 10% FBS), medium 2 (TCM-199 + 10% FBS with eCG [10 IU/ml] and hCG [5 IU/ml]), or medium 3 (TCM-199 + 10% FBS with eCG, hCG, and 17beta-estradiol [2 microg/ml]). After 24 h of maturation in vitro, the maturation rate of oocytes cultured in medium 2 (70%, n = 79) was significantly greater (P < 0.01) than those of oocytes cultured in the other two media (27%-36%, n = 67-73). At 48 h, similar maturation rates (56%-69%, n = 76-87) were observed for all three types of media. For activation experiments, oocytes cultured in medium 2 were stimulated with electrical and chemical stimuli either individually or in combination. Treatment with cycloheximide and 6-dimethylaminopurine (6-DMAP) following electrical stimulation resulted in 43% (n = 58) of the oocytes developing to the blastocyst stage. Such an activation rate represented a significant improvement over those obtainable under other tested conditions, including individual treatment with electrical pulses (10%, n = 41), cycloheximide (3%, n = 58), or 6-DMAP (5%, n = 59). Blastocysts derived from in vitro activation appeared to be normal morphologically and were composed of an appropriate number of both inner cell mass (mean +/- SEM, 10.3 +/- 1.1; n = 11) and trophectoderm (60.8 +/- 2.9, n = 11) cells. These results have begun to elucidate parameters important for animal modeling and cloning with ferrets.     

影响猪体细胞核移植重构胚体外发育的若干因素

以卵丘细胞为核供体细胞组成重构胚,卵裂率达到56.7%,发育至桑椹胚达11.7%、孵化囊胚率为6.7%,显著高于成纤维细胞组成的重构胚(p<0.05)。我们研究了卵母细胞的采集方法,激活方法和卵龄对卵丘细胞核移植重构胚体外发育的影响。以血清饥饿法将卵丘细胞诱导至GO或G1期,抽吸法/解剖法采集卵母细胞,体外培养33或44 h,将卵丘细胞置于去核卵母细胞的卵周隙中,重构胚以钙离子载体A23817或电脉冲结合6-DMAP激活处理,体外培养6天,结果表明,卵母细胞采集方法、激活液中细胞松弛素(CB)并不影响重构胚的发育(以卵龄44h的卵母细胞为受体);而以电脉冲结合6-DMAP激活处理能提高重构胚发育能力(以卵龄33 h的卵母细胞为受体)(p<0.05)。本研究显示,以电脉冲结合6-DMAP激活卵丘细胞重构胚,能在体外发育至囊胚     

银鲫卵母细胞体外诱导成熟技术的建立

研究以银鲫为材料, 根据银鲫(Carassius auratus gibelio)卵母细胞生发泡(Germinal vesicle, GV)边移程度及剥离GV中减数分裂前期染色体的凝集状态, 将银鲫Ⅳ时相的卵母细胞分为GV0、GV1、GV2和GV3四个时期; 并进一步比较了分别处于这4个时期银鲫卵母细胞体外诱导培养的成熟率、卵裂率和孵化率。结果表明, GV1期之后的卵母细胞均可有效进行体外诱导成熟, 可正常受精发育, 由于GV1期卵母细胞有较长时间用于显微操作, 因此GV1期卵母细胞被选为进行体外诱导的最早时期的卵母细胞。以GV1期卵母细胞为研究材料, 摸索了银鲫卵母细胞体外诱导成熟的适宜条件: 取GV1期的Ⅳ时相卵母细胞, 放置于pH 8.5、加有1 μg/mL孕酮激素(17α, 20β-dihydroxy-4-pregnen-3-one, DHP)的格氏平衡盐溶液(Gey’s balanced salt solution, GBSS)中, 在23℃培养箱中体外诱导12h后, 将滤泡膜剥离后再进行人工体外授精, 其所获胚胎的孵化率可达55.5%。此外, 将体外转录合成的带GFP标签的h2af1o mRNA注射到GV1期卵母细胞, 发现经显微操作和体外诱导后不仅可以通过GFP绿色荧光信号活体观察GVBD、受精、卵裂和早期胚胎发育的全过程, 而且诱导成熟的卵子仍可正常受精和胚胎发育。研究建立的银鲫卵母细胞体外诱导成熟技术为银鲫和其他鱼类卵母细胞发育过程研究及其相关基因和细胞显微操作提供了技术平台。     

Effect of co-culture with theca interna on nuclear maturation of horse oocytes with low meiotic competence,and subsequent fusion and activation rates after nuclear transfer

We conducted this study to examine whether or not co-culture with theca cells improves the maturation rate of horse oocytes with compact cumuli and to evaluate the cytoplasmic competence of oocytes after maturation by assessing fusion, activation and cleavage rates after nuclear transfer. We collected oocytes by scraping follicles from slaughterhouse-derived ovaries and classified them as having an expanded or a compact cumulus. Expanded oocytes were matured in M199 supplemented with 10% FBS and 5 microU/ml FSH for 24 h: compact oocytes were cultured in the same medium, or they were co-cultured in the same medium with theca interna explants, for 24 or 42 h. Oocytes were held with or without 10 microg/ml cytochalasin B, before washing and micromanipulation. and they were fused with donor fibroblasts by electrical pulse. Fused oocytes were activated with Ca ionophore/cycloheximide, cultured for 5 days, and stained with Hoechst to assess nuclear development. We considered oocytes with an enlarged nucleus, or having cleavage with multiple nuclei, to be activated. There was no significant difference in overall maturation rate between compact oocytes cultured with theca and compact controls. When these two groups were combined, there was a significant increase in the proportion of oocytes in MII between 24 and 42 h (P < 0.05). Expanded oocytes had a significantly higher rate of maturation than did compact oocytes (64% versus 25-30%; P < 0.001). There were no significant differences in rates of successful enucleation, fusion, activation or cleavage between compact control and compact + theca oocytes, nor between compact and expanded oocytes; however, expanded oocytes treated with cytochalasin B had a significantly higher survival rate after enucleation than did untreated expanded oocytes (P < 0.05). Three embryos developed from recombined oocytes, with maximum cleavage to 10 cells. The results of this study indicate that co-culture with theca cells does not increase either nuclear or cytoplasmic maturation of compact oocytes. Cytochalasin B is helpful in increasing survival of horse oocytes during enucleation. In vitro matured equine oocytes have the potential to develop into embryos after nuclear transfer; this is the first full report of production of cloned embryos in this species.     

The effect of harvesting technique on efficiency of oocyte collection and different maturation media on the nuclear maturation of oocytes in camels (Camelus dromedarius)

The purpose of this investigation was to develop an efficient method for harvesting oocytes from dromedary camel ovaries and to examine the effect of different maturation media on their subsequent maturation in vitro. Oocytes were collected by aspirating the follicular contents using a needle attached to a syringe (Method I, n=163 ovaries) or to a constant aspirating pressure, applied by a vacuum pump (Method II, n=117 ovaries). Individual follicles were excised from ovaries and follicles were punctured with two needles (Method III, n=117). Oocytes were matured in vitro for 40-42 h. At the end of maturation period, oocytes were denuded of cumulus cells and the proportion of oocytes in metaphase-II (MII) stage was determined. In the second experiment, oocytes collected by the dissection method were matured in Tissue Culture Medium199 (TCM), CR1 or modified Connaught Medical Research Laboratories medium-1066 (CMRL) and their nuclear maturation was evaluated after 40-42 h. The recovery rate of oocytes was higher (P<0.01) with Method III compared with Method I or II (94, 31 and 33%, respectively). A higher proportions of oocytes collected with Method I or II were either completely or partially denuded compared with Method III (31, 14% versus 1%). The proportions of viable oocytes (78, 60 and 70%, respectively) and those showing metaphase II was not different (39, 50 and 46%, respectively, P>0.05) among the three treatment groups. Oocyte maturation rate was higher (P<0.05) when TCM was used compared with CMRL or CR1 medium. There was, however, no difference in the maturation rate for oocytes cultured in CMRL or CR1 medium. It may be concluded that a higher proportion of cumulus enclosed oocytes may be recovered by follicle dissection method compared to aspiration using syringe or pump. The higher recovery rate with a comparable proportion of viable and matured oocytes resulted in the overall increase in the number of matured (MII) oocytes/ovary with follicle dissection procedure compared with aspiration procedures. For in vitro maturation of oocytes, TCM is superior to CR1 and CMRL as basic maturation medium for this species.