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Nghiên cứu ảnh hưởng của một số yếu tố kỹ thuật sử dụng tinh trùng đã xác định tính biệt cái đến khả năng sản xuất phôi in vitro và in vivo trên bò Holstein Friesian (Luận văn thạc sĩ)Nghiên cứu ảnh hưởng của một số yếu tố kỹ thuật sử dụng tinh trùng đã xác định tính biệt cái đến khả năng sản xuất phôi in vitro và in vivo trên bò Holstein Friesian (Luận văn thạc sĩ)Nghiên cứu ảnh hưởng của một số yếu tố kỹ thuật sử dụng tinh trùng đã xác định tính biệt cái đến khả năng sản xuất phôi in vitro và in vivo trên bò Holstein Friesian (Luận văn thạc sĩ)Nghiên cứu ảnh hưởng của một số yếu tố kỹ thuật sử dụng tinh trùng đã xác định tính biệt cái đến khả năng sản xuất phôi in vitro và in vivo trên bò Holstein Friesian (Luận văn thạc sĩ)Nghiên cứu ảnh hưởng của một số yếu tố kỹ thuật sử dụng tinh trùng đã xác định tính biệt cái đến khả năng sản xuất phôi in vitro và in vivo trên bò Holstein Friesian (Luận văn thạc sĩ)Nghiên cứu ảnh hưởng của một số yếu tố kỹ thuật sử dụng tinh trùng đã xác định tính biệt cái đến khả năng sản xuất phôi in vitro và in vivo trên bò Holstein Friesian (Luận văn thạc sĩ)Nghiên cứu ảnh hưởng của một số yếu tố kỹ thuật sử dụng tinh trùng đã xác định tính biệt cái đến khả năng sản xuất phôi in vitro và in vivo trên bò Holstein Friesian (Luận văn thạc sĩ)Nghiên cứu ảnh hưởng của một số yếu tố kỹ thuật sử dụng tinh trùng đã xác định tính biệt cái đến khả năng sản xuất phôi in vitro và in vivo trên bò Holstein Friesian (Luận văn thạc sĩ)
IH IH NGUY N TH TUY T NHUNG U Y U T K THU T S D N KH NG C A M T S NH N XU HOLSTEIN FRIESIAN LU KHOA H P - 2015 IH IH NGUY N TH TUY T NHUNG U Y U T K THU T S D N KH NG C A M T S NH N XU : 60.62.01.05 LU C P ng d n khoa h c: m TS - 2015 i ii Sinh iii i ii iii vi vii viii 1.1 B 1.1.1 1.1.3 1.2.1 .7 1.3.1 , .10 12 12 13 1.4.3 16 17 18 iv .20 .21 22 24 25 27 27 27 .27 27 itro .27 28 28 28 29 29 32 .34 35 35 3.1.1 3.1.2 Ch 35 37 v 39 .41 45 .45 47 48 50 .51 3.3.2 53 53 55 56 57 3.4.3 58 61 62 68 vi BSA : Bovine Serum Albumin FCS : Foetal Calf serum FSH : Follicle Stimulating Hormone GnRH : Gonadotropin Releasing Hormone HEPES : N-(2 Hydroxyethyl) piperazine- HF : Holstein Friesian LH : Luteinizing Hormone LTH : Luteo Tropic Hormone PRH : Prolactin Releasing Hormone MEM : Minimum Essential Medium PGF : Prostaglandin F SOF : Synthetic oviduct fluid TCM-199 : Tisue culture medium 199 PCR : Polymerase chain reaction NST : -(2-ethanesulforic acid) vii 3.1: c 35 37 39 42 45 47 c 49 51 53 54 p 56 57 59 viii h 1.1: 11 17 23 33 36 38 39 40 42 : .43 7: 44 50 : 52 3.10: 55 57 60 60 nang 2 61 in vitro (P < 0,05) (P < 0,05) tinh (P < 0,05) in vitro -T - /ml in vivo 62 , , , in vitro Lan (2010), , (4), tr 36 39 -702 , , tr 140-143 , , tr 117-118 in vitro , , 75 82 Adams G P., Matteri R, L,, Ginther O J (1992), Effect of progesterone on ovarian follicles, emergence of follicular waves and circulating follicle-stimulating hormone in heifers , Journal of Reproduction and Fertility, 95, pp 627-640 Ahmad N., Townsend E C., Dailey R A., Inskeep E.K (1997), Relationship of hormonal patterns and fertility to occurrence of two or three waves of ovarian follicles, before and after breeding, in beef cows and heifers , Animal Reproduction Science, 49, pp 13-28 Bavister B D (1996), Culture of preimplantation embryos: facts and artifacts , Hum Reprod Update, 1, pp 91-148 Bearden H J., Fuquay J W (2000), The female reproduction system , Applied Animal reproduction, Prentice Hall, InC, pp 7-20 63 10 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embryos by a visual colorimetric assay of the X lingked enzyme, glucose-6-phosphat dehydrogenase , Theriogenology, 25, pp 733 55 Wilson R D., Fricke P M., Leibfried-Rutledge M L., Rutledge J J., Syverson C M., Wwigel K A (2006), In vitro production of bovine embryo using sex-sorted sperm Theriogenology, 65, pp 1007-1015 56 Zhang M., Lu K H., Seidel G E (2003), Development of bovine embryos after in vitro fertilization of oocytes with flow cytometrically sorted, stained and unsorted sperm , Theriogenology, 60, pp:1657 1663 57 Ziebe S., Petersen K., Lindenberg S., Anderson A G., Gabrielsen A., Andersen A N (1997), Embryo morphology or cleavage stage: How to select the best embryo for transfer after in vitro fertilization , Human Reproduction, 12, pp 1545-1549 68 - - - - Sanyo CO2 Incubator - -325 2000 - Pippete pasteur ainer disk, Nunc disk 69 - - Sucrose C12H22O11 196-00015 Wako Japan Disodium Hydrogenphosphate Na2HPO4 197-02865 Wako Japan D(+) Glucose C6H12O6 041-00595 Wako Japan Potassium Dihydrogenphosphate KH2PO4 165-04242 Wako Japan Potassium Chloride KCl 163-03545 Wako Japan Sodium Hydrogen Carbonate NaHCO3 191-01305 Wako Japan Sodium Chloride NaCl 191-01665 Wako Japan Sodium Dihydrogen Phosphate Dihydrate NaH2CO3.2H2O 192-02815 Wako Japan Calcium Chloride Dihydrate CaCl2.2H2O 031-00435 Wako Japan Phenol Red Solution 0,5% P0290 Sigma Magnesium Chloride Hexahydrate MgCl2.6H2O 131-00162 Wako Japan Ethylene Glycol C2H6O2 Prolabo Germany 70 Glycerol Prolabo Germany Medium 199 3769 GIBCO BME Amino Acide Solution 50x B-6766 Sigma MEM Non-Essential Amino Acide Solution 100x 02380 GIBCO Penicilin G Potassium 200.000 Meiji Seika Kaisha LTD Tokyo Japan Novo-Heparin Pharmaceutical Products LTD V ventis Japan Streptomycin Sulphate 1g Meiji Kaisna LTD Tokyo Japan L-Glutamic Acid G-4815 Sigma Albumin, Bovine A-4378 Sigma Caffein Sodium Benzoate C-4144 Sigma L(+)-Lactic Acid L-4388 Sigma Albumin, Bovine A-7030 Sigma Sodium Pyruvate 199-03062 Wako Japan - PGF2 - Estradiol -) NaCl 8,0g 71 KCl 0,2g Na2HPO4 1,15g KH2PO4 0,2g 700ml CaCl2 0,1g MgCl2.6H2O 0,1g 200ml - penicilin/ml + 100 l streptomycin/ml 100.000iu penicilin/ml + 100 l streptomycin/ml TCM 199 47,5 ml CS 2,5 ml Penicilin-G 10.000IU (100IU/ml) Streptomycin Sulfate 72 NaCl 4,3092g KCl 0,1974g CaCl2.2H2O 0,2171g NaH2PO4.2H2O 0,0840g MgCl2.6H2O 0,0697g 0,1ml NaHCO3 2,5873g Phenol Red 0,5% 0,04ml 76ml Na pyruvate 0,01375g Penicilin 10.000IU Streptomycin 10mg 24ml 50ml Na Caffein benzoate 0,1942g 73 Heparin (Novo-heparin: 5000IU/ml) 40ml BSA 400mg 10ml BSA 200mg NaCl 6,7031g KCl 0,2311g Na Pyruvate 0,0440g NaHCO3 2,2011g Phenol Red 5% 2ml Hemicalcium lactate 0,5996g 76ml B 20ml BME Essential Amino Acids 2ml MEM Non Essential Amino Acids 1ml 74 L-Glutamic Acid (2mg/ml) 1ml BSA 3mg/ml Penicilin-G 10.000IU (100IU/ml) Streptomycin Sulfate - VIGRO Freeze medium ... 3.10: 55 57 60 1 , nhu C in , vivo, in vitro c Song, sinh , h , in vivo in vitro in vitro in vivo in vitro vivo - in 1.1 B 1.1.1 sinh, ( ( ) ) , 0,3g, 10 - 20g , , , 15 mm... vitro in vitro, in vitro in vitro in vitro ( 1988) [49] , , in vitro in vivo 18 , in vitro in vitro (Bavister, 1996) [8] Tuy , Defined Medium ( 1975) [11] in vitro , 20 in vitro , 1.5 C, CO2 19 ,... in vitro (Sales v , 2009) [10] , 2011) [42] 1.4 DNA DNA DNA, DNA DNA DNA (Ganner , 2006) [17], s , 17 g Theo (Johnson and Welch 1999) [26] 1.5 in vitro in vitro in vitro in vitro, in vitro in