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1 * * The Effects of Oocyte Oocytes Intracytoplasmic ICSI) Kee TaekHoo Hai * SikChun Department of Obstetrics and Gynecology, School of Medicine, Kyungpook Taegu, *Department of Animal Science, Taegu Kyungbuk, Korea Objective: ICSI, cumulus and corona cells should be removed from the oocytes hyaluronidase) and mechanical (pipetting) methods But little is known about the effects of different degrees of oocytedenudation ICSI The aim of this study was to evaluate the effects of varying the degrees of oocytedenudation ICSI Methods: oocytes) were grouped into group A and B according to the degree of denudation, complete and partial, respectively In experiment 2, patients (oocytes) were grouped into group, and according to the length of incubation time of denuded oocytes 2 and >2 hours, respectively Results: oocytedenudation ICSI oocytes ICSI, survival rates was higher in group (831%) than in group (615%, P<005) or group (643%) However no statistically significant differences were found between incubation time and fertilization or development rates Conclusions: ICSI oocytedenudation However, the denuded oocytes ICSI Key Words: oocytedenudation, Incubation time, Intracytoplasmic 1998

2 , Palermo (1) ICSI), ICSI ICSI,, 26 hyaluronidase pipette ICSI 78ICSI corona cell cumulus cell hyaluronidase pipetting,78 pipette m 10IU 7 Veld(8) ICSI incubation ( ) ICSI M, ICSI incubation ICSI ICSI, incubation

3 1 F-10 Nutrient Mixture Medium (Ham's F-10, , Gibco, USA)12 g/ NaHCO3 ICSI Dulbecco's Modified Eagle Medium (DMEM, , Gibco, USA) Vero monolayer Tissue Culture Medium 199 (TCM-199, , Gibco, USA) 05% antibiotics (Streptomycin sulfate, S-9137; Penicillin -G, P-3032, Sigma, USA), (Osmomat 030, Gonatec, Germany) 280 mos/ 02 (Millex GV, Millipore, USA) 14 tube (2001, Falcon, USA) 4 95%, 375% CO2 (3154, Forma, USA) 6 2 gonadotrophin releasing hormone agonist (GnRH-a) Mid-luteal phase 900 Buserelin (Suprefact, Hoechst, Germany), estradiol human menopausal gonadotropin (hmg, Pergonal, Serono, Italy) follicle stimulating hormone (FSH, Urofollotropin, Metrodin, Serono, Italy) ,000 IU human chorionic gonadotrophin (hcg, Profasi, Serono, Italy), hcg % CO2 (IVF chamber, Hoffman, USA) (SMZ-10, Nicon, Japan) 01% hyaluronidase 2 Ham's F-10 ICSI ICSI 1 2 1) 1: ICSI (ICSI, ), ICSI 2 (Group A) (Group B) ICSI (,, ) ( ) 2) 2: ICSI, 1 (Group ), 12 (Group ) 2 (Group ) ICSI 3 (hff, human follicular fluid) hff hff (3,500 rpm)2(30, 10) (ICSI) Holding pipette (P-2174, Sigma, USA) , 10-15,

4 sperm injection pipette (ICSI micropipette, Humagen, USA) 7-8, 5-6 DPBS3% polyvinylpyrolidone (PVP, PVP-360, Sigma, USA)10 paraffin oil (294365H, BDH, UK) ICSI ( 1 )DMEM (10% hff), PVP injection pipette injection pipette , injection pipette activation 2-3, 37 5% CO2 5 Vero monolayer Vero Ouhibi (9), cell flask 4 ( ), trypsin cell suspension3 flask,, monolayer 2 200,000 cells 3 Vero Ouhibi (9) 6 Vero monolayer Vero 20% hff DMEM, 2-3

5 7 ICSI, SAS package (10) t-test 5%

6 1 ICSI ICSI (ICSI, ) ICSI 2 (Group A) (Group B) ICSI, Table 1 (Group A) (Group B) ICSI, (773% : 792%), (2 PN; 659% : 542%) (/2PN; 983% : 962%) group A (659%)group B (542%) 2 ICSI ICSI ICSI ICSI, 1 (Group ), 12 (Group ) 2 (Group ) ICSI (Table 2) ICSI, group (831%)group (615%) group (643%) (Group : Group ; P<005, Group : Group ; NS), (2PN; 462% : 500% : 593%) (667% : 100% : 100%)

7 ICSI,,,, 24,68,11,12 ICSI 13,14 sperm parameter ICSI 5,14 Ca2+ ICSI 16,17, Yanagida (18)ICSIelectrical activation 100%, ICSI Palermo (6) ICSI immobilization, proteinlipid, decondensation immobilization, Chen (19) immobilization mid-piece Chen (19) immobilization Tasdemir (20) ICSI 400, 9% Palermo (6) (oolemma) (sudden breakage),, ( ) Conaghan (21) 2-4 EBSS pyruvate-047mm, glucose-free medium (22) glutamine DMEM M DMEM (hff) (23) ICSI hyaluronidase pipetting 78 (7) 10, IU/, ICSI ICSI, pipette ICSI, (unknown harmful effect) (10 IU/) 1000 pipette 01% (66 IU/), pipette ,, pipette ICSI hyaluronidase pipetting ICSI pathenogenetic activation,24,25 Van de Veld (8) ICSI incubation ICSI incubation ICSI ICSI, ICSI 2

8 ICSI ICSI,, (2 PN) 2 (659%) (542%) ICSI ICSI ICSI Van de Veld (8) ICSI incubation ( ) ICSI M incubation, (<1, 1-2, >2 ) ICSI, (<1 : 1-2; P<005, 1-2 : >2; NS) (2PN) 2, ICSI, ICSI ICSI 2 ICSI

9 1 Palermo G, Joris H, Devroey P, Van Steirteghem AC Pregnancies after intracytoplasmic injection of a single spermatozoon into an oocyte Lancet Hum Reprod Hum Reprod Hum Reprod Hum Reprod Hum Reprod Hum Reprod Hum Reprod Hum Reprod Fertil Steril Fertil Steril Hum Reprod Hum Reprod Fertil Steril Fertil Steril Hum Reprod Hum Reprod

10 Hum Reprod Hum Reprod in vittro J Reprod Fertil,,, 1998; 25: ,,,,, (Human Body Fluids) 1994; 21: Palermo G, Joris H, Derde MP et al: Sperm characteristics and outcome of human assisted fertilization by subzonal insemination and intracytoplasmic sperm injection Fertil Steril 1993; 59: Van Steirteghem AC, Joris H, Liu J Protocol for intracytoplasmic sperm injection Hum Reprod

11 Group A Group B Complete denudation Partial denudation No of cycles 12 9 No of injected oocytes No of survived oocytes (%) 68 (773) 38 (792) No of 2 PN status oocytes (%) 58 (659) 26 (542) No of cleaved embryos / 2 PN (%) 57 (983) 25 (962) No significant difference between group A and B

12 Table 2 Effects of incubation period (< 1, 12 or > 2 hours) from denudation to sperm injection on the outcomes of ICSI Group Group Group < 1 hr 12 hrs > 2 hrs No of cycles No of injected oocytes No of survived oocytes (%) 24 (615)* 9 (643) 49 (831)* No of 2 PN status oocytes (%) 18 (462) 7 (500) 35 (593) No of cultured embryos No of cleaved embryos (%) 10 (667) 7 (100) 31 (100) *

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1.. Volume 12, Number 1, 6~16, Factors influencing consultation time and waiting time of ambulatory patients in a tertiary teaching hospital Jee-In Hwang College of Nursing Science, Kyung Hee University :

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