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1.
为研究不同剂量的IGF-1和EGF对辽宁绒山羊毛囊体外生长及毛囊形态的影响,采用显微分离法分离绒山羊初级毛囊,分别测定不同浓度IGF-1(0.1、1、10和100 ng/mL)、不同浓度EGF(0.2、2、20和100 ng/mL)、联合添加IGF-1和EGF(分别添加10 ng/mL和20 ng/mL)对毛囊生长和形态变化的影响.结果表明,IGF-1和EGF对毛囊体外生长均具有正向调节作用,作用大小与剂量有关,最适宜浓度IGF-1为10 ng/mL;EGF为20 ng/mL.IGF-1(10ng/mL)和EGF(20 ng/mL)联合添加对毛囊生长促进作用优于单独添加.IGF-1刺激毛乳头和毛母质细胞分裂增殖;EGF促进毛囊外根鞘细胞分化;联合添加刺激毛球部增大.  相似文献   

2.
生长因子对山羊无腔卵泡卵母细胞体外生长的影响   总被引:10,自引:0,他引:10  
山羊无腔卵泡卵母细胞在胰岛素样生长因子I(IGF-I)和表皮生长因子(EGF)及碱性成纤维细胞生长因子(bFGF)单独或联合存在时,与FSH-道体外培养9d,结果表明,IGF-I(100ng/mL)能有效地维持卵泡卵母细胞的存活并促进其生长,存活率达75.00%,相对生长率达45.06%;EGF(50ng/mL)能提高卵泡卵母细胞的体外存活率,类似于IGF-I,但对其生长相对表现出抑制影响;bFGF(50ng/mL)对卵泡卵母细胞的体外存活具有促进作用,但对卵母细胞生长的促进作用不明显(卵母细胞的存活率为76.92%,相对生长率为31.55%)。IGF-I(100ng/mL)和EGF(50ng/mL)联合存在时,不论是对卵泡卵母细胞的存活,还是对生长均产生了最好的效果,卵母细胞存活率达到90.91%,相对生长达48.24%。本试验结果一个共同点是在FSH存在的情况下,这3种生长因子单独或有选择的联合,对山羊卵泡卵母细胞的体外存活能力均具有正向调节作用。  相似文献   

3.
在卵母细胞体外成熟液(含FSH)中分别添加10, 30, 50ng/mL表皮生长因子(EGF),24h检查其成熟率.结果表明:添加 EGF50ng/mL时牛卵母细胞第一极体排出率显著提高,达85.4%(P<0.01);同时发现在含FSH和EGF的成熟液中添加HMG(尿促性素)并不能提高牛卵母细胞体外成熟效果(P>0.05);同时还发现单独添加HMG成熟效果显著好于FSH(P<0.05).  相似文献   

4.
The study investigated the effects of epidermal growth factor(EGF) and insulin-like growth factor 1(IGF-I),alone or together,on the in vitro maturation and cleavage of ovine oocytes,aimed to optimize the in vitro maturation conditions for ovine oocytes.The results showed that the maturation and cleavage rates were 71.2% and 45.5% respectively when the medium was supplemented with 50 ng/mL EGF alone,which was significantly higher than other EGF supplemented groups (0,10,20,30,and 40 ng/mL) (P<0.05).The highest maturation and cleavage rates were 72.9% and 45.7% when the EGF concentration reached 100 ng/mL.The maturation and cleavage rates were 70.7% and 58.5% with 40 ng/mL IGF-I supplemented,which were significantly higher than other treatments (0,10,20,60,80,and 100 ng/mL) (P<0.05).The lowest maturation and cleavage rates were 38.8% and 20.0% when the IGF-I concentration reached 100 ng/mL (P<0.05).When 50 ng/mL EGF and 40 ng/mL IGF-I were used concomitantly,the maturation and cleavage rates were 85.6% and 61.0% respectively,which were significantly higher than the treatments with EGF or IGF-I alone (P<0.05).  相似文献   

5.
为了分析外源激素促卵泡激素(FSH)对绵羊卵丘颗粒细胞增殖的影响,试验分离并体外培养了绵羊卵丘颗粒细胞,然后使用促卵泡激素受体(FSHR)免疫组织化学法鉴定了分离培养的细胞,进而分析了添加不同浓度FSH对细胞增殖速度的影响。结果表明:将卵丘颗粒细胞从卵丘颗粒细胞-卵母细胞复合体(COCs)上分离24 h之后,能够观察到部分卵丘颗粒细胞开始贴壁,传代之后细胞形态未发生明显变化。FSHR免疫组织化学法鉴定结果证明分离培养的绵羊卵丘颗粒细胞能够特异性地表达FSHR。培养基中添加FSH后,在培养前4 d,高浓度(100 ng/mL)和低浓度(10 ng/mL)的FSH均能促进卵丘颗粒细胞的增殖,但在培养6 d之后,高浓度的FSH对卵丘颗粒细胞的增殖存在抑制作用。说明外源激素FSH在不同浓度下对卵母颗粒细胞的增殖具有不同的影响。  相似文献   

6.
通过建立和完善绵羊子宫内膜上皮原代细胞体外培养技术,为研究绵羊母体和孕体之间的相互作用机制建立体外着床模型。采用组织块法分离培养子宫内膜上皮细胞,观察其生长情况,并比较不同表皮生长因子(EGF)浓度对子宫内膜上皮细胞增殖的作用效果。结果显示,组织块培养1~2 d后,组织周围迁出子宫内膜上皮细胞,长满60 mm培养皿需9~12 d;经差时消化法纯化后,F1代绵羊子宫内膜上皮细胞纯度可达90%以上,表明所获细胞可用于后续试验;F2代细胞在不同浓度EGF(0、12.5、25、50、75、100 ng/mL)下培养144、168 h,经检测在144 h,12.5、25和100 ng/mL浓度下D_(450 nm)值极显著高于对照组(P0.01),50 ng/mL浓度下显著高于对照组(P0.05),在168 h,12.5 ng/mL浓度下显著高于对照组(P0.05),因此,在12.5 ng/mL EGF作用下效果最佳;F3代细胞在0、12.5 ng/mL浓度下培养不同时间(24、48、72、96、120、144、168、192、216 h),经检测D_(450 nm)值在144 h差异显著(P0.05),168 h后差异极显著(P0.01)。因此,在培养液中添加12.5 ng/mL EGF可以更加高效、快速得到子宫内膜上皮细胞。  相似文献   

7.
表皮生长因子对水牛胚胎体外发育及凋亡的影响   总被引:1,自引:0,他引:1  
为了探讨表皮生长因子(EGF)对水牛早期胚胎体外发育及凋亡的影响,通过收集屠宰场卵巢卵母细胞进行体外成熟和体外受精,将假定的受精卵置于含不同浓度EGF(0,25,50和100 ng/mL)的培养液中培养,检查分裂率和囊胚发育率,用细胞凋亡试剂盒(Annexin-V-FluosStaining kit)试剂染色,统计囊胚细胞凋亡率和坏死率。结果表明:50 ng/mL EGF组的孵化囊胚率显著高于对照组(P<0.05),该组细胞凋亡率和坏死率显著低于对照组(P<0.05)。100 ng/mL EGF的卵裂率、囊胚率、D7囊胚率和孵化囊胚率显著低于对照组和其他试验组(P<0.05)。细胞凋亡率和坏死率显著高于其他各组(P<0.05)。提示:一定浓度的EGF可提高囊胚孵化率,并可抑制胚胎细胞的凋亡。  相似文献   

8.
试验旨在研究不同激素配比及表皮生长因子(EGF)浓度对牛卵母细胞体外成熟及卵母细胞质量的影响。将随机分组的卵丘-卵母细胞复合体于添加FSH+LH、HMG、FSH+LH+E2、HMG+E2 4种不同激素组合配比的成熟基础液中培养,对比其体外成熟率,比较了EGF对牛卵母细胞体外成熟率和孤雌胚胎体外发育的影响,并采用TUNEL法检测添加不同浓度EGF的牛孤雌激活囊胚细胞凋亡情况。结果表明,添加HMG的成熟试验结果稳定,E2对牛卵母细胞成熟有一定的促进作用,HMG+E2联合使用可以得到高效稳定的成熟结果;在此基础上,在成熟液中添加30 ng/mL EGF对牛卵母细胞的成熟质量、胚胎发育及降低胚胎细胞凋亡都有明显的促进作用。因此,在体外成熟培养液中添加0.075 IU/mL HMG、1 μg/mL E2和30 ng/mL EGF对牛卵母细胞的成熟和质量较为有益。  相似文献   

9.
为研究表皮生长因子(EGF)、β-巯基乙醇(β-ME)、亚硫磺酸(HTAU)对牛卵母细胞体外成熟(IVM)及孤雌激活胚胎体外发育(IVC)效果的影响,实验采集牛卵巢采用切割法收集卵丘-卵母细胞复合体(COCs)随机处于不同浓度EGF、β-ME培养液成熟培养24 h后孤雌激活,研究其在不同胚胎培养液中的后续发育,以期筛选出最好的牛卵母细胞IVM-IVC条件。结果表明:添加25、50、100 ng/mL EGF组的成熟率、卵裂率均高于对照组,其中50 ng/mL EGF组的成熟率、卵裂率与对照组差异显著(P<0.05);50、100、500μmol/L的β-ME对牛卵母细胞体外成熟没有促进作用;50 ng/mL EGF与50、100μmol/Lβ-ME组合并无协同作用;胚胎培养基中添加不同浓度EGF对早期胚胎的体外发育无显著影响;而添加100μmol/Lβ-ME组的囊胚率、囊胚细胞数均极显著高于对照组(P<0.01);在成熟液中添加50 ng/mL EGF和100μmol/Lβ-ME、胚胎培养液中添加0.5 mmol/L HTAU孤雌胚发育最好。  相似文献   

10.
本试验旨在研究表皮生长因子(EGF)对脂多糖(LPS)刺激的应激状态下仔猪肠道钠磷转运载体蛋白Ⅱb(NaPi-Ⅱb)表达的影响。本试验由2部分组成,1)细胞试验:以猪小肠上皮细胞(IPEC-J2)为模型,试验设4个组,对照组(0 ng/mL EGF,0μg/mL LPS)、EGF组(100 ng/mL EGF,0μg/mL LPS)、LPS组(0 ng/mL EGF,1.0μg/mL LPS)、EGF+LPS组(100 ng/mL EGF,1.0μg/mL LPS),每组3个重复; 2)动物试验:选取24头体重接近、健康状况良好的21日龄"长白×大白"二元杂交断奶阉公猪[平均体重(5.76±0.38) kg],随机分为4个组,对照组(基础饲粮)、EGF组(基础饲粮+2 mg/kg EGF)、LPS组(基础饲粮+腹腔注射100μg/kg BW LPS)、EGF+LPS组(基础饲粮+2 mg/kg EGF+腹腔注射100μg/kg BW LPS),每组6个重复,每个重复1头猪。结果表明:1)细胞试验中,与对照组相比,EGF组IPEC-J2细胞NaPi-Ⅱb mRNA及蛋白表达显著下降(P0.05),而EGF+LPS组细胞NaPi-Ⅱb mRNA及蛋白表达显著增加(P0.05),EGF与LPS免疫应激的互作效应对NaPi-Ⅱb mRNA及蛋白表达影响显著(P0.05)。2)动物试验中,各组间血清钙(Ca)含量无显著差异(P0.05),LPS组血清磷(P)含量显著高于对照组、EGF组、EGF+LPS组(P0.05),EGF组血清碱性磷酸酶(ALP)活性显著高于LPS组(P 0.05),EGF与LPS免疫应激互作效应对血清P含量影响显著(P 0.05),对血清Ca含量和ALP活性影响不显著(P 0.05)。与对照组相比,EGF组空肠与回肠NaPi-Ⅱb mRNA表达显著下降(P 0.05),而EGF+LPS组空肠与回肠NaPi-Ⅱb mRNA表达显著增加(P0.05),EGF与LPS免疫应激互作效应对空肠与回肠NaPi-Ⅱb mRNA表达影响显著(P0.05)。细胞与动物试验结果表明,EGF对NaPi-Ⅱb的表达起抑制作用,但在免疫应激状态下可促进NaPi-Ⅱb的表达,表明EGF可促进应激状态下肠道P的主动转运。  相似文献   

11.
The object of this study was to investigate the role of epidermal growth factor (EGF) and IGF-I in the regulation of preantral follicular growth, antrum formation, and granulosal cell proliferation/ apoptosis. Porcine preantral follicles were manually dissected and cultured for up to 8 d in Waymouth's (Exp. 1) or alpha-minimum Eagle's essential medium (Exp. 2 and 3) supplemented with 10 microg/mL of transferrin, 100 microg/mL of L-ascorbic acid, and 2 mU/mL of ovine FSH, in the presence (Exp. 1 and 3) or absence (Exp. 2) of 7.5% fetal calf serum. According to the experimental protocol, IGF-I (0, 1, 10, or 100 ng/mL; Exp. 1), or IGF-I (50 ng/mL), EGF (10 ng/mL) and EGF+IGF-I (Exp. 2 and 3) were added to the culture media. In Exp. 1, follicles exhibited a concentration-dependent response (P < 0.05) to IGF-I, with the highest rates of granulosal cell proliferation, follicular integrity, and recovery rate of cumulus cell-oocyte complexes and lowest incidence of apoptosis occurring at the highest IGF-I dose. In Exp. 2 serum-free medium, granulosal cell proliferation was low (1 to 5%), irrespective of whether EGF and/or IGF-I were present and cellular apoptosis was increased (P < 0.05) on d 4 and 8 in the EGF+IGF-I group compared with the addition of either factor alone. In Exp. 3, granulosal cell proliferation was high in all follicles cultured in serum-containing medium for the first 3 d, but fell sharply (P < 0.05) on d 4, except in media containing IGF-I. Collectively, EGF and IGF-I increased granulosal cell proliferation, decreased apoptosis, and promoted follicular antrum formation. These results may provide useful information for developing a preantral follicular culture system in which the oocytes are capable of fertilization and embryonic development.  相似文献   

12.
为观察表皮生长因子 ( epidermalgrowth factor,EGF)对成年水牛晶状体上皮细胞增殖作用的影响。将不同浓度 EGF作用于体外培养的成年水牛晶状体上皮细胞 ,采用 MTT法测定细胞的增殖能力。结果 :EGF在浓度为 1ng/ m L和 1 0 ng/ m L作用的前 3 d促增殖作用逐渐加强 ,呈现时间依赖性 ,且在第 3 d达到最大促增殖效果。不同浓度的 EGF对晶状体上皮细胞的增殖作用不同 ,浓度在 1 0 ng/ m L作用 2 4h后即有明显的促增殖作用 ( P <0 .0 5) ,而作用72 h后最低的有效浓度为 1 ng/ m L,而且 2 50ng/ m L EGF有最大促增殖作用。结论 :EGF是诱导成年水牛晶状体上皮细胞增殖的重要因素。  相似文献   

13.
The objectives of the present studies were 1) to develop a culture system that has the positive effect of serum on granulosa cell attachment and allows subsequent expression of hormonal effects in serum-free medium and 2) to determine the effect of insulin, epidermal growth factor (EGF), estradiol (E2), and growth hormone (GH) on growth, steroidogenesis, and(or) protein synthesis of bovine granulosa cells. Cells from small (1 to 5 mm) and large (greater than 8 mm) follicles were collected from cattle and cultured for either 4 or 6 d. When cells from small follicles were cultured, insulin (5 micrograms/ml) increased (P less than .05) cell numbers (cells x 10(5)/well) severalfold compared with controls. Alone, EGF (10 ng/ml), FSH (200 ng/ml), LH (200 ng/ml), E2 (2 micrograms/ml), or GH (0 to 1,000 ng/ml) had no effect on cell numbers. However, when included with insulin, 30, 100, and 300 ng/ml of GH increased (P less than .05) granulosa cell numbers on d 4 of culture. Insulin alone increased (P less than .05) progesterone production (ng.10(5) cells-1.24 h-1) by severalfold on d 4, but EGF, FSH, LH, or GH alone had no effect and E2 inhibited progesterone production. In the presence of insulin, FSH and GH (100 ng/ml) increased (P less than .05) progesterone production on d 4 of culture, whereas EGF (10 ng/ml) elicited a decrease (P less than .05) in production. In cells from both sizes of follicles, GH (300 ng/ml) increased synthesis of cellular proteins (greater than 10 kDa). In cells from only large follicles, LH (200 ng/ml) decreased synthesis and secretion of proteins (greater than or equal to 3.5 kDa). These results support the hypothesis that GH may have direct effects on bovine ovarian function.  相似文献   

14.
【目的】探究表皮生长因子(epidermal growth factor,EGF)和成纤维细胞生长因子2(fibroblast growth factor 2,FGF-2)对猪皮下脂肪神经嵴干细胞(neural crest stem cells,NCSCs)增殖及分化的影响,以优化猪皮下脂肪神经嵴干细胞的培养条件。【方法】通过体外分离培养原代猪皮下脂肪NCSCs,免疫荧光染色鉴定NCSCs标志物p75 NTR,并用不同浓度的EGF和FGF-2(0和0、10和10、10和20、20和10、20和20、30和30 ng/mL)作用于传代猪皮下脂肪NCSCs,用CCK-8试剂盒测定细胞增殖率,确定细胞生长的最适EGF和FGF-2浓度,将试验分为空白组和最适浓度组,测定两组细胞的生长曲线,成脂化诱导后油红O染色,对比两组细胞的脂滴生成量。【结果】免疫荧光染色结果显示,原代猪皮下脂肪NCSCs经p75 NTR鉴定呈阳性。CCK-8细胞增殖试验结果显示,EGF和FGF-2的浓度均为20 ng/mL时对传代猪皮下脂肪NCSCs的促增殖作用最佳。生长曲线显示,两组细胞均在第5~9天处于对数生长期,第10~15天细胞增殖减缓,逐渐到达停滞期。油红O染色结果显示,最适浓度组胞质内的脂滴生成量远多于对照组。【结论】在培养液中添加20 ng/mL EGF和20 ng/mL FGF-2对猪皮下脂肪NCSCs的增殖和分化有促进作用。  相似文献   

15.
瘦素和FSH对绵羊卵泡颗粒细胞孕激素分泌的影响   总被引:1,自引:0,他引:1  
试验旨在探明瘦素和卵泡刺激素(FSH)对绵羊卵泡颗粒细胞孕激素分泌的影响并建立瘦素和FSH之间的相互作用。从屠宰场收集健康母羊的卵巢,机械分离卵泡,收集卵泡颗粒细胞。在细胞培养液中加入不同浓度的FSH(0 U/mL,2.5 U/mL,5 U/mL,7.5 U/mL,10 U/mL)培养24 h和48 h,通过MTT检测细胞增殖,以确定后续试验FSH的使用浓度。在细胞培养液中加入瘦素(0 ng/mL,25 ng/mL,50 ng/mL)和FSH(5 U/mL)单独或联合作用48 h后,通过ELISA检测细胞培养液中孕酮(P_4)的浓度。收集细胞,通过qPCR和Western blotting检测CYP11A1、STAR、3β-HSD的表达。结果表明:5 U/mL的FSH浓度可显著促进细胞生长,使类固醇相关基因(CYP11A1、STAR、3β-HSD)的表达升高(P<0.05),但对其蛋白无显著影响,对P4分泌无刺激作用。单独添加瘦素时,25 ng/mL瘦素降低了CYP11A1、STAR、3β-HSD基因的表达(P<0.05),但相应蛋白无显著性差异,同时对P_4的分泌无影响;50 ng/mL瘦素降低了STAR、CYP11A1基因的表达,但相应蛋白表达和P4分泌未受影响。瘦素和FSH联合使用时,P4分泌显著降低,STAR表达降低(P<0.05),但对其他类固醇合成基因和蛋白(CYP11A1、3β-HSD)的表达无显著影响(P>0.05)。综上所述,FSH对绵羊卵泡颗粒细胞增殖和类固醇合成基因表达有促进作用;瘦素对FSH诱导下P_4的分泌有抑制作用,且具有剂量依赖性,提示瘦素参与卵泡的发育过程。  相似文献   

16.
To determine the effect of gonadotropins on insulin- and insulin-like growth factor (IGF-I)-induced bovine granulosa cell functions, granulosa cells from bovine ovarian follicles were cultured for 2 days in the presence of 10% fetal calf serum (FCS), and then cultured for an additional 2 days in serum-free medium with added hormones. In the presence of 0 or 1 ng/mL of insulin or IGF-I, FSH had little or no effect (P>0.05) on estradiol production by granulosa cells from both small (1–5 mm) and large (≥8 mm) follicles. However, in the presence of ≥3 ng/mL of insulin, FSH increased (P<0.05) estradiol production by granulosa cells from small and large follicles such that the estimated dose (ED50) of insulin necessary to stimulate 50% of the maximum estradiol production was decreased by 2- to 3-fold from 22 to 28 ng/mL in the absence of FSH to 7–14 ng/mL in the presence of FSH. Similarly, in the presence of ≥3 ng/mL of IGF-I, FSH increased (P<0.05) estradiol production by granulosa cells from small and large follicles such that the ED50 of IGF-I for estradiol production was decreased by 4- to 5-fold from 25 to 36 ng/mL in the absence of FSH to 5–6 ng/mL in the presence of FSH. In the presence of FSH, the maximal effect of insulin on estradiol production was much greater than that of IGF-I (137- versus 12-fold increase) and were not additive; when combined, 100 ng/mL of IGF-I completely blocked the stimulatory effect of 100 ng/mL of insulin. In the absence of FSH, the maximal effect of insulin and IGF-I on estradiol production was similar. Concomitant treatment with 30 ng/mL of LH reduced (P<0.05) insulin-stimulated estradiol production by 52% on day 1 and 19% on day 2 of treatment. Insulin, IGF-I and FSH also increased (P<0.05) granulosa cell numbers and progesterone production but their maximal effects were less (i.e., <4-fold increase) than their effects on estradiol production. In conclusion, insulin and IGF-I synergize with FSH to directly regulate ovarian follicular function in cattle, particularly granulosa cell aromatase activity.  相似文献   

17.
Objective The physiologic mechanisms involving growth factors, including PDGF‐BB, EGF, and TGF‐β1, as potent mediators of fibroblasts and epithelial cells in corneal wound healing remain unknown. The goal of this study was to determine culture methods for equine epithelial cells and keratocytes and to investigate how exogenous growth factors influence proliferation of both cell types. Procedures Cell cultures were established from healthy corneas harvested from horses immediately following euthanasia and maintained using standard tissue culture protocols. To determine the effects of PDGF‐BB, EGF, TGF‐β1, keratocytes (1 × 105/well) and epithelial cells (2 × 105/well) were each cultured in 12 well plates and exposed separately to the growth factors. The cells were exposed to concentrations of EGF between 0 and 50 ng/mL; PDGF‐BB between 0 and 75 ng/mL; and TGF‐β1 between 0 and 10 ng/mL. Cell proliferation was measured using 3H‐thymidine assay and differences in growth determined using anova and Tukey's HSD test (P < 0.05). Results Epithelial cell and keratocyte cultures were successfully established. EGF maximally stimulated keratocyte and epithelial cells at 25 ng/mL and 5 ng/mL, respectively. PDGF‐BB maximally stimulated keratocytes and epithelial cells at 50 ng/mL and 5 ng/mL, respectively. TGF‐β1 inhibited keratocytes at 5 ng/mL and 10 ng/mL, and epithelial cells at 1 ng/mL and 2 ng/mL. Conclusions Methods were established to maintain epithelial cells and keratocytes in vitro. PDGF‐BB and EGF stimulate, while TGF‐β1 inhibits the proliferation of epithelial cells and keratocytes. These growth factors may play a role in maintenance and repair of the equine cornea.  相似文献   

18.
Goat preantral follicles were cultured to investigate the effects of insulin-like growth factor-I (IGF-I), epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) on the in vitro growth and viability of oocytes. Preantral follicles were isolated mechanically and enzymatically (using collagenase and DNase) from prepuberal goat ovaries. The working medium was composed of Defined Eagle's Minimum Essential Medium (DMEM) supplemented with HEPES (20 mM), 10% fetal calf serum (FCS), hypoxanthine (2 mM), dibutyryl cyclic adenosine 3',5'-monophosphate (dbcAMP) (2 mM), penicillin (75 ng/ml) and streptomycin (50 ng/ml). The culture medium consisted of the working medium with follicle stimulating hormone (FSH) (100 ng/ml) and hydrocortisone (40 ng/ml) added. In the experiment, goat preantral follicles were cultured for 9 days in the culture medium and in the culture medium supplemented with either IGF-I (100 ng/ml), EGF (50 ng/ml), bFGF (50 ng/ml) or IGF-I (100 ng/ml)+EGF (50 ng/ml). The results indicated that IGF-I (100 ng/ml) effectively maintained the survival of oocytes and promoted their growth; EGF (50 ng/ml) enhanced the survival rate of oocytes but had a negative effect on oocyte growth; bFGF (50 ng/ml) stimulated oocyte survival but had no obvious effect on their growth while IGF-I (100 ng/ml) and EGF (50 ng/ml) in combination had a greater effect on both survival and growth rate of oocytes than IGF-I or EGF alone. The supplementation of IGF-1 and EGF to the culture medium is recommended in the culture of goat preantral follicles.  相似文献   

19.
通过建立和完善绵羊子宫内膜上皮原代细胞体外培养技术,为研究绵羊母体和孕体之间的相互作用机制建立体外着床模型。采用组织块法分离培养子宫内膜上皮细胞,观察其生长情况,并比较不同表皮生长因子(EGF)浓度对子宫内膜上皮细胞增殖的作用效果。结果显示,组织块培养1~2 d后,组织周围迁出子宫内膜上皮细胞,长满60 mm培养皿需9~12 d;经差时消化法纯化后,F1代绵羊子宫内膜上皮细胞纯度可达90%以上,表明所获细胞可用于后续试验;F2代细胞在不同浓度EGF(0、12.5、25、50、75、100 ng/mL)下培养144、168 h,经检测在144 h,12.5、25和100 ng/mL浓度下D450 nm值极显著高于对照组(P<0.01),50 ng/mL浓度下显著高于对照组(P<0.05),在168 h,12.5 ng/mL浓度下显著高于对照组(P<0.05),因此,在12.5 ng/mL EGF作用下效果最佳;F3代细胞在0、12.5 ng/mL浓度下培养不同时间(24、48、72、96、120、144、168、192、216 h),经检测D450 nm值在144 h差异显著(P<0.05),168 h后差异极显著(P<0.01)。因此,在培养液中添加12.5 ng/mL EGF可以更加高效、快速得到子宫内膜上皮细胞。  相似文献   

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