In vitro development of one-cell embryos from outbred mice: Influence of culture medium composition

1990 ◽  
Vol 26 (2) ◽  
pp. 151-156 ◽  
Author(s):  
Akiko Spindle
Keyword(s):  
Culture Medium ◽  
Medium Composition ◽  
In Vitro Development ◽  
Outbred Mice ◽  
Vitro Development

Effect of LH and Estradiol-17β Supplementation at Different Time Points on In vitro Development of Preantral Follicles in Sheep

2020 ◽  
Author(s):  
L.S.S. Varaprasad Reddy ◽  
B.R. Naik ◽  
A.V.N. Sivakumar ◽  
B. Punyakumari ◽  
J. Suresh
Keyword(s):  
In Vitro Maturation ◽  
Culture Medium ◽  
Follicular Development ◽  
Standard Medium ◽  
In Vitro Development ◽  
Preantral Follicles ◽  
Time Points ◽  
Vitro Development ◽  
Estradiol 17Β

Background: Ovarian follicular development and growth are controlled by many hormones and growth factors. Despite the fact that LH and estradiol-17β have been utilized for the in vitro culture of preantral follicles yet, the suitable time points of supplementation of LH and estradiol-17β is not known. Therefore this study aimed to investigate the influence of addition of LH and estradiol-17β at different time points on in vitro development of preantral follicles (PFs’) in sheep. Method: Preantral follicles isolated from the ovarian cortical slices using micro dissection method were cultured for six days in Bicarbonate buffered Tissue culture medium 199B (TCM 199B) or in a standard culture medium supplemented with LH (2 μg/ml) and estradiol-17β (5 ng/ml) at different points during the culture period. COCs isolated from the follicles at the end of six day culture in different treatments were subjected to in vitro maturation for additional 24h. Result: Supplementation of LH and estradiol-17β during last two days of the culture supported better proportion of PFs’ exhibiting growth whereas supplementation of LH and estradiol-17β during first two days of the culture supported better average increase in diameter and proportion of PFs’ exhibiting antrum formation at the end of six day culture. Further the oocytes in COCs isolated at the end of culture in these treatments and subsequently subjected to in vitro maturation (IVM) for 24hr developed at a higher frequency to MII (metaphase II) stage. Supplementation of LH and estradiol-17β to TCM 199B culture medium in early stages followed by standard medium alone in later stages supports better development of PFs’ in vitro. Following supplementation with LH and estradiol-17β for the first two days culture of PFs’ in standard medium appears to be advantageous for the development of preantral follicles in vitro.

Bovine dominant follicular fluid promotes the in vitro development of goat preantral follicles

2012 ◽  
Vol 24 (3) ◽  
pp. 490 ◽  
Author(s):  
A. B. G. Duarte ◽  
V. R. Araújo ◽  
R. N. Chaves ◽  
G. M. Silva ◽  
D. M. Magalhães-Padilha ◽  
...  
Keyword(s):  
Follicular Fluid ◽  
Culture Medium ◽  
Follicular Development ◽  
Ultrastructural Analysis ◽  
In Vitro Development ◽  
Preantral Follicles ◽  
Nuclear Membranes ◽  
Vitro Development ◽  
First Time

The aim of this study was to evaluate the effect of follicular fluid collected from bovine dominant follicles (bFF) on the in vitro development of goat preantral follicles and determine the best time to add this supplement to the culture medium. The preantral follicles were isolated and randomly distributed into four treatments in absence (control) or presence of 10% of bFF added on Days 0 (FF0–18), 6 (FF6–18) or 12 (FF12–18) of culture onwards. After 18 days, follicular development was assessed based on follicular survival, antral cavity formation, increased follicular diameter as well as fully grown oocyte (>110 μm) viability and meiosis resumption. The oocytes from the cultured follicles were in vitro-matured and processed for fluorescence or ultrastructural analysis. The results showed that on Day 18 the treatment FF0–18 had a significantly higher (P < 0.05) survival than control and FF12–18, but not FF6–18. The addition of bFF at the beginning of culture (FF0–18 and FF6–18) promoted a high percentage of follicular growth, meiosis resumption and early antrum formation. Moreover, this study described for the first time the ultrastructural analysis of caprine oocytes grown in vitro. This evaluation revealed that in the presence of bFF on (FF0–18) the in vitro-grown oocytes presented normal organelle distribution and well-defined, intact plasma and nuclear membranes. In conclusion, bFF originating from dominant follicles maintain the survival and promote the in vitro growth of goat preantral follicles when added at the beginning of culture.

In vitro development of theDrosophila chorion in a chemically defined organ culture medium

1979 ◽  
Vol 186 (4) ◽  
pp. 351-362 ◽  
Author(s):  
William H. Petri ◽  
Mihalis N. Mindrinos ◽  
Mary F. Lombard ◽  
Lukas H. Margaritis
Keyword(s):  
Organ Culture ◽  
Culture Medium ◽  
In Vitro Development ◽  
Organ Culture Medium ◽  
Vitro Development

Glucose and glycine synergistically enhance the in vitro development of porcine blastocysts in a chemically defined medium

2012 ◽  
Vol 24 (3) ◽  
pp. 443 ◽  
Author(s):  
Tomomi Mito ◽  
Koji Yoshioka ◽  
Shoko Yamashita ◽  
Chie Suzuki ◽  
Michiko Noguchi ◽  
...  
Keyword(s):  
Culture Medium ◽  
Survival Rates ◽  
Defined Medium ◽  
Blastocyst Stage ◽  
Atp Content ◽  
Blastocyst Development ◽  
In Vitro Development ◽  
Defined Culture ◽  
Vitro Development

In the present study, the effects of glucose and/or glycine on the in vitro development of Day 5 (Day 0 = IVF) porcine blastocysts were determined. The addition of 2.5–10 mM glucose to the chemically defined culture medium porcine zygote medium (PZM)-5 significantly increased blastocyst survival rates compared with those of blastocysts cultured in the absence of glucose. The addition of 5 and 10 mM glycine to PZM-5 containing 5 mM glucose significantly enhanced the development to hatching and the number of hatched blastocysts compared with no addition of glycine. However, the addition of glycine to PZM-5 with no glucose did not improve blastocyst development. The ATP content of Day 6 blastocysts cultured with glucose was significantly higher than that of blastocysts cultured in the absence of glucose, regardless of glycine supplementation. The diameter and total cell numbers were significantly greater, and the apoptotic index was significantly lower, in Day 6 blastocysts cultured with both glucose and glycine. These results indicate that glucose is an important energy source for the porcine blastocyst and that glucose and glycine act synergistically to enhance development to the hatching and hatched blastocyst stage in vitro.

Effect of culture medium replacement protocol on the in vitro development of isolated caprine secondary follicles

2011 ◽  
Vol 95 (2-3) ◽  
pp. 139-143 ◽  
Author(s):  
Valdevane Rocha Araújo ◽  
Roberta Nogueira Chaves ◽  
Ana Beatriz Graça Duarte ◽  
Juliana Jales de Hollanda Celestino ◽  
Gerlane Modesto da Silva ◽  
...  
Keyword(s):  
Culture Medium ◽  
In Vitro Development ◽  
Vitro Development

Effect of RU486 on different stages of mouse preimplantation embryos in vitro

1990 ◽  
Vol 68 (11) ◽  
pp. 1457-1460 ◽  
Author(s):  
Subhash C. Juneja ◽  
Melvin G. Dodson
Keyword(s):  
Embryo Development ◽  
Culture Medium ◽  
Inhibitory Effect ◽  
Blastocyst Stage ◽  
Preimplantation Embryos ◽  
In Vitro Development ◽  
Stage Embryo ◽  
Morula Stage ◽  
Vitro Development

17β-Hydroxy-11β-(4-dimethylaminophenyl)-17α-(1-propynyl)estra-4,9-dien-3-one (RU486) inhibited the in vitro development of different stages of mouse preimplantation embryos under study. Two-celled embryos, morulae, and early blastocysts were obtained from B6D2F1 mice. The embryos were grown in Ham F-10 nutrient mixture (with glutamine) supplemented with sodium bicarbonate (2.1 g/L), calcium lactate (282 mg/L), and bovine serum albumin (fraction V, 3 mg/mL) at 37 °C in a humidified incubator supplied with 5% CO2 in air. RU486 was added to the culture medium at concentrations of 1, 5, 10, and 20 μg/mL. Culture medium with 0.05% ethanol served as the control. In vitro growth of embryos was assessed by the following criteria: (i) two-celled stage embryo development to blastocyst stage after 72 h, (ii) morula stage grown to blastocyst stage after 24 h, and (iii) early blastocyst stage development to hatching blastocyst after 12 h, in culture. RU486 inhibited the in vitro development of two-celled embryos, morulae, and early blastocysts at concentrations of 5, 10, and 20 μg/mL culture medium (p < 0.001). The inhibitory effect of RU486 at these concentrations on the development of all the stages of embryos under study was irreversible. However, RU486 did not affect embryo development at 1 μg/mL culture medium. The study indicates the direct adverse effect of RU486 at 5 μg/mL and higher concentrations in culture medium on the development of mouse preimplantation embryos in vitro, and it encourages its further investigation as a postcoital contraceptive in animal models and humans.Key words: RU486, mouse, preimplantation embryos, embryo culture, postcoital contraceptive.

Sign in / Sign up

Export Citation Format

Share Document