Effect of chemical or electrical activation of bovine oocytes on blastocyst development and quality.
Activation of in vitro-matured (IVM) oocytes is essential for successful embryo production following nuclear transfer (NT) or intracytoplasmic sperm injection (ICSI). This study was designed to compare the rates of blastocyst production and embryo quality (as measured by numbers of viable cells) following parthenogenetic activation with electrical pulse or the use of two different calcium ionophores, A23187 (CA) or ionomycin (IO), with or without the addition of bovine serum albumin (BSA). IVM oocytes with a first polar body were randomly allocated to five treatment groups: CA (5 microM CA, 5 min; n = 88), CA + BSA (5 microM CA, 5 min; BSA, 5 min; n = 90), IO (5 microM IO, 5 min; n = 91), IO + BSA (5 microM IO, 5 min; BSA, 5 min; n = 86) and EL (two pulses of 1.5 kV/cm, 20 micros; n = 120). Blastocyst rates were higher (p < 0.05) for CA (54.4%), IO (51.4%) and EL (54.5%) than for IO + BSA (18.3%). Treatment CA + BSA (39.8%) did not differ from the others. There was no difference (p > 0.05) among treatments in total number of cells. However, the percentage of viable cells was reduced in CA (49.9%), CA + BSA (45.8%), IO (64.9%), IO + BSA (50.9%) compared with EL (82.7%). In summary, the addition of BSA to the IO treatment had an adverse effect on blastocyst production rates. Although there was no difference between electrical stimulation and chemical activation on blastocyst production rates, electrical activation resulted in blastocysts with a higher percentage of viable cells.
Milazzotto
M P
MP
Department of Animal Reproduction, Faculty of Veterinary Medicine, Laboratory of In Vitro Fertilization, Cloning and Animal Transgenesis, University of Sao Paulo, Sao Paulo, Brazil.
Feitosa
W B
WB
Department of Animal Reproduction, Faculty of Veterinary Medicine, Laboratory of In Vitro Fertilization, Cloning and Animal Transgenesis, University of Sao Paulo, Sao Paulo, Brazil.
Coutinho
Ars
A
Department of Animal Reproduction, Faculty of Veterinary Medicine, Laboratory of In Vitro Fertilization, Cloning and Animal Transgenesis, University of Sao Paulo, Sao Paulo, Brazil.
Goissis
M D
MD
Department of Animal Reproduction, Faculty of Veterinary Medicine, Laboratory of In Vitro Fertilization, Cloning and Animal Transgenesis, University of Sao Paulo, Sao Paulo, Brazil.
Oliveira
V P
VP
Department of Animal Reproduction, Faculty of Veterinary Medicine, Laboratory of In Vitro Fertilization, Cloning and Animal Transgenesis, University of Sao Paulo, Sao Paulo, Brazil.
Assumpção
Meoa
M
Department of Animal Reproduction, Faculty of Veterinary Medicine, Laboratory of In Vitro Fertilization, Cloning and Animal Transgenesis, University of Sao Paulo, Sao Paulo, Brazil.
Visintin
J A
JA
Department of Animal Reproduction, Faculty of Veterinary Medicine, Laboratory of In Vitro Fertilization, Cloning and Animal Transgenesis, University of Sao Paulo, Sao Paulo, Brazil.
D000818:Animals; D001755:Blastocyst / Q000187:drug effects / Q000502:physiology*; D002417:Cattle / Q000196:embryology*; D002452:Cell Count / Q000662:veterinary; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D019976:Cloning, Organism; D004305:Dose-Response Relationship, Drug; D004558:Electric Stimulation / Q000379:methods*; D004622:Embryo, Mammalian / Q000502:physiology*; D005260:Female; D053652:Nuclear Transfer Techniques / Q000662:veterinary; D009865:Oocytes / Q000187:drug effects / Q000502:physiology*; D010312:Parthenogenesis / Q000502:physiology; D012710:Serum Albumin, Bovine / Q000009:adverse effects / Q000494:pharmacology*; D020554:Sperm Injections, Intracytoplasmic / Q000662:veterinary; D013997:Time Factors
D012710:Serum Albumin, Bovine