Effect of maturation medium supplements on bovine oocyte fertilization and embryo development

Theriogenology

193

EFFECT OF MATURATION MEDIUM SUPPLEMENTS ON BOVINE OOCYTE FERTILIZATION AND EMBRYO DEVELOPMENT C. MacCallum, D. Salamone and A.T. Palasz Department of Herd Medicine and Theriogenology, WCVM, University of Saskatchewan, Saskatoon, Saskatchewan, 5B4 OWO, Canada. An experiment was designed to determine the benefit of serum or serum substitutes and oFSH on in vitro development of bovine oocytes and embryos. Cumulus oocyte complexes (COC) were collected from slaughterhouse ovaries and within 4 to 5 h were placed in maturation media. A total of 587 COC were placed randomly into 1 of 6 groups in 6 replicates: 1) control-TCM-199 with 0.25 mm01 Na pyruvate and antibiotics, 2) as control+200 rig/ml oFSH, 3) as control+O. 1% ET surfactant (ETS; Vetrephamr Can. Ic), 4) as control+0 l%ETS+2OO @ml oFSH, 5) as control+5% FCS and 6) as control+fi% FCS and 200 r&ml oFSH. Twelve to 20 COC were placed in 400 ul of the respective maturation media under paraffin oil and cultured at 39 “C and 5%CO2 in humidified air. After 23 h, COC were inseminated (IVF) with 1x107/ml t?ozen-thawed sperm washed twice with BrdckettOliphant (BO) medium supplemented with 5mM caffeine and 20 ug/ml heparin. After 6 h, the presumptive zygotes were placed into 500 ul drops of TCM-199+10% FCS (12 to 20 per drop under paraffin oil) and co-cultured with cumulus cells at 39 “C and 5% CO2 in humidihed air. Cleavage rates were evaluated on Day 2 (insemination = Day 0) and the numbers of blastocysts were counted on Days 7. 8, and 9. and compared by Chi-square analysis. Results are shown in table 1. Table 1. Cleavage and embryo development rates from oocytes matured in TCM- 199 plus fetal calf serum (FCS) or ET surf&ant (ETS) in presence or absence of oFSH n

Cleaved (%)

Morulae (%)

Blastocyst (%)

1) Control

105

51(49) il

35(33) il

13(12) il

2) TCM- 199+oFSH

112

77(69) hc

45(40) il

17(15) il

3) TCM-199+ETS

7-l

50(65) hc

29(38) ’

7(9) Cl

53(7 Dhc

34(45) d

lO( 13) il

Group

4) TCM- 199+ETS+oFSH 5) TCM- 199+FCS

1 IO

83(75) hc

43(39) ?’

6(5) h

6) TCM- 199+FCS+oFSH

108

86(8(j) ”

46(43) i’

17(16) il

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