In vitro maturation of bitch oocytes from advanced preantral follicles in synthetic oviduct fluid medium: serum is not essential

The ability of oocytes from preantral follicles to mature in vitro was assessed using a synthetic oviduct fluid (SOF) medium. Advanced preantral follicles (approximately 210μm diameter) were isolated from the ovaries of domestic bitches and assigned to one of four treatment groups: (1) SOF (n=230);...

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Published inTheriogenology Vol. 58; no. 9; pp. 1689 - 1703
Main Authors Bolamba, Digbo, Russ, Kara D, Olson, Mary A, Sandler, Jody L, Durrant, Barbara S
Format Journal Article
LanguageEnglish
Published United States Elsevier Inc 01.12.2002
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Abstract The ability of oocytes from preantral follicles to mature in vitro was assessed using a synthetic oviduct fluid (SOF) medium. Advanced preantral follicles (approximately 210μm diameter) were isolated from the ovaries of domestic bitches and assigned to one of four treatment groups: (1) SOF (n=230); (2) SOF+3mg/ml bovine serum albumin (+BSA, n=220); (3) SOF+20% fetal bovine serum (+FBS, n=227); or (4) SOF+3mg/ml BSA+20% FBS (+BSA+FBS, n=232), then cultured for up to 72h. A group of control follicles was not cultured (n=103). The percentages of oocytes reaching metaphase I to metaphase II stages (MI to MII) did not differ between treatments at each culture period. Within treatments, the percentages of oocytes at MI to MII stages did not differ with duration of culture. However, when compared to the control group (0.97%) the percentages of oocytes at MI to MII increased (P<0.05) in the SOF group after 48h (10.0%) and 72h (12.2%) of culture. In the +BSA (10.1%) and +FBS (9.7%) groups, the percentages of oocytes at MI to MII increased (P<0.05) above control values only after 72h of culture. The percentage of oocytes at MI to MII did not significantly increase in the +BSA+FBS group (3.9, 6.6 and 7.6% at 24, 48 and 72h of culture, respectively) compared to the control group. These results indicate that under the described conditions supplementation of culture medium with BSA or FBS is not essential, and the simple medium SOF can support nuclear maturation of a small proportion of bitch oocytes in vitro.
AbstractList The ability of oocytes from preantral follicles to mature in vitro was assessed using a synthetic oviduct fluid (SOF) medium. Advanced preantral follicles (approximately 210 microm diameter) were isolated from the ovaries of domestic bitches and assigned to one of four treatment groups: (1) SOF (n = 230); (2) SOF + 3 mg/ml bovine serum albumin (+BSA, n = 220); (3) SOF + 20% fetal bovine serum (+FBS, n = 227); or (4) SOF + 3 mg/ml BSA + 20% FBS (+BSA+FBS, n = 232), then cultured for up to 72 h. A group of control follicles was not cultured (n = 103). The percentages of oocytes reaching metaphase I to metaphase II stages (MI to MII) did not differ between treatments at each culture period. Within treatments, the percentages of oocytes at MI to MII stages did not differ with duration of culture. However, when compared to the control group (0.97%) the percentages of oocytes at MI to MII increased (P &lt; 0.05) in the SOF group after 48 h (10.0%) and 72 h (12.2%) of culture. In the +BSA (10.1%) and +FBS (9.7%) groups, the percentages of oocytes at MI to MII increased (P &lt; 0.05) above control values only after 72 h of culture. The percentage of oocytes at MI to MII did not significantly increase in the +BSA+FBS group (3.9,6.6 and 7.6% at 24,48 and 72 h of culture, respectively) compared to the control group. These results indicate that under the described conditions supplementation of culture medium with BSA or FBS is not essential, and the simple medium SOF can support nuclear maturation of a small proportion of bitch oocytes in vitro.
The ability of oocytes from preantral follicles to mature in vitro was assessed using a synthetic oviduct fluid (SOF) medium. Advanced preantral follicles (approximately 210μm diameter) were isolated from the ovaries of domestic bitches and assigned to one of four treatment groups: (1) SOF (n=230); (2) SOF+3mg/ml bovine serum albumin (+BSA, n=220); (3) SOF+20% fetal bovine serum (+FBS, n=227); or (4) SOF+3mg/ml BSA+20% FBS (+BSA+FBS, n=232), then cultured for up to 72h. A group of control follicles was not cultured (n=103). The percentages of oocytes reaching metaphase I to metaphase II stages (MI to MII) did not differ between treatments at each culture period. Within treatments, the percentages of oocytes at MI to MII stages did not differ with duration of culture. However, when compared to the control group (0.97%) the percentages of oocytes at MI to MII increased (P<0.05) in the SOF group after 48h (10.0%) and 72h (12.2%) of culture. In the +BSA (10.1%) and +FBS (9.7%) groups, the percentages of oocytes at MI to MII increased (P<0.05) above control values only after 72h of culture. The percentage of oocytes at MI to MII did not significantly increase in the +BSA+FBS group (3.9, 6.6 and 7.6% at 24, 48 and 72h of culture, respectively) compared to the control group. These results indicate that under the described conditions supplementation of culture medium with BSA or FBS is not essential, and the simple medium SOF can support nuclear maturation of a small proportion of bitch oocytes in vitro.
The ability of oocytes from preantral follicles to mature in vitro was assessed using a synthetic oviduct fluid (SOF) medium. Advanced preantral follicles (approximately 210 microm diameter) were isolated from the ovaries of domestic bitches and assigned to one of four treatment groups: (1) SOF (n = 230); (2) SOF + 3 mg/ml bovine serum albumin (+BSA, n = 220); (3) SOF + 20% fetal bovine serum (+FBS, n = 227); or (4) SOF + 3 mg/ml BSA + 20% FBS (+BSA+FBS, n = 232), then cultured for up to 72 h. A group of control follicles was not cultured (n = 103). The percentages of oocytes reaching metaphase I to metaphase II stages (MI to MII) did not differ between treatments at each culture period. Within treatments, the percentages of oocytes at MI to MII stages did not differ with duration of culture. However, when compared to the control group (0.97%) the percentages of oocytes at MI to MII increased (P < 0.05) in the SOF group after 48 h (10.0%) and 72 h (12.2%) of culture. In the +BSA (10.1%) and +FBS (9.7%) groups, the percentages of oocytes at MI to MII increased (P < 0.05) above control values only after 72 h of culture. The percentage of oocytes at MI to MII did not significantly increase in the +BSA+FBS group (3.9,6.6 and 7.6% at 24,48 and 72 h of culture, respectively) compared to the control group. These results indicate that under the described conditions supplementation of culture medium with BSA or FBS is not essential, and the simple medium SOF can support nuclear maturation of a small proportion of bitch oocytes in vitro.
Author Durrant, Barbara S
Russ, Kara D
Bolamba, Digbo
Olson, Mary A
Sandler, Jody L
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  organization: Reproductive Physiology Division, Center for Reproduction of Endangered Species, Zoological Society of San Diego, P.O. Box 120551, San Diego, CA 92112-0551, USA
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Keywords Meiosis
Preantral follicle
Oocyte
Reproductive technology
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Snippet The ability of oocytes from preantral follicles to mature in vitro was assessed using a synthetic oviduct fluid (SOF) medium. Advanced preantral follicles...
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pubmed
elsevier
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StartPage 1689
SubjectTerms Animals
Body Fluids
Cells, Cultured
Culture Media
Culture Media, Serum-Free
Dogs
Fallopian Tubes - metabolism
Female
Meiosis
Metaphase
Oocyte
Oocytes - physiology
Oocytes - ultrastructure
Ovarian Follicle - cytology
Preantral follicle
Reproductive technology
Serum Albumin, Bovine
Title In vitro maturation of bitch oocytes from advanced preantral follicles in synthetic oviduct fluid medium: serum is not essential
URI https://dx.doi.org/10.1016/S0093-691X(02)01080-4
https://www.ncbi.nlm.nih.gov/pubmed/12472139
https://search.proquest.com/docview/72759232
Volume 58
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