Rat embryo development in vitro

1981 ◽  
Vol 96 (4) ◽  
pp. 546-551 ◽  
Author(s):  
S. K. Roy ◽  
Jayasree Sengupta ◽  
S. K. Manchanda
Keyword(s):  
Bovine Serum Albumin ◽  
Bovine Serum ◽  
Culture Medium ◽  
Synthetic Medium ◽  
Blastocyst Formation ◽  
Rat Embryo ◽  
Rat Embryos ◽  
Morula Stage ◽  
Development In Vitro

Abstract. We report the first successful culture of 8-cell/morula stage rat embryos in a fully synthetic medium supplemented with 3% crystalline bovine serum albumin. Eighty-four per cent of morulae developed to blastocysts, showing that this is a highly efficient culture medium for in vitro studies on rat pre-implantation embryos. Blastocyst formation was severely inhibited by antioestrogen (nafoxidine 3 μg/ml) but no further reversal was obtained by giving oestrogen to culture medium containing this antagonist.

Effect of serum on the in vitro maturation of canine oocytes

1999 ◽  
Vol 11 (8) ◽  
pp. 387 ◽  
Author(s):  
Takeshige Otoi ◽  
Maya Fujii ◽  
Aya Ooka ◽  
Masaki Tanaka ◽  
Tatsuyuki Suzuki
Keyword(s):  
Bovine Serum Albumin ◽  
Bovine Serum ◽  
In Vitro Maturation ◽  
Culture Medium ◽  
The Other ◽  
Serum Supplement ◽  
Meiotic Competence ◽  
Positive Effect ◽  
Serum Type

The meiotic competence of canine oocytes cultured for 72 h in medium supplemented with three different concentrations (5, 10 and 20%) of anoestrous, oestrous or metoestrous bitch serum, or with 0.3% bovine serum albumin (BSA), was examined. The oestrous serum supplement had a positive effect on the resumption of meiosis, compared with the other supplements (P<0.05). The number of oocytes that reached metaphase I (MI) and metaphase II (MII) was significantly higher (P<0.05) with the oestrous serum supplement than with the anoestrous serum supplement. There were no significant differences among the three different concentrations in each serum type with respect to the proportion of oocytes that completed meiosis (MI to MII). The number of oocytes that resumed meiosis in the 10% oestrous serum supplement was significantly higher (P<0.05) than those of each concentration of the anoestrous and metoestrous serum supplements, and of the 0.3% BSA supplement. Moreover, a higher number of oocytes reached MII in the presence of the 10% oestrous serum supplement than with the 10% anoestrous serum supplement. These results suggest that supplementation of the culture medium with 10% oestrous serum is the optimal treatment for in vitro maturation of canine oocytes.

scholarly journals PS48 can replace bovine serum albumin in pig embryo culture medium, and improve in vitro embryo development by phosphorylating AKT

2015 ◽  
Vol 82 (4) ◽  
pp. 315-320 ◽  
Author(s):  
Lee D. Spate ◽  
Alana Brown ◽  
Bethany K. Redel ◽  
Kristin M. Whitworth ◽  
Randall S. Prather
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Embryo Development ◽  
Embryo Culture ◽  
Bovine Serum ◽  
Culture Medium ◽  
Embryo Culture Medium

scholarly journals 144 ROLE OF GnRH ON MOUSE PRE-IMPLANTATION EMBRYONIC DEVELOPMENT IN VITRO

2005 ◽  
Vol 17 (2) ◽  
pp. 222
Author(s):  
M. Montagner ◽  
A. Cropp ◽  
J. Swanson ◽  
R. Cederberg ◽  
P. Goncalves ◽  
...  
Keyword(s):  
Embryonic Development ◽  
Gnrh Agonist ◽  
Critical Role ◽  
Blastocyst Stage ◽  
Dependent Manner ◽  
Blastocyst Formation ◽  
Morula Stage ◽  
Development In Vitro ◽  
Human Uterine Endometrium

The interaction between GnRH and its receptor on gonadotropes within the anterior pituitary gland represents a key point for regulation of the reproduction. In addition, GnRH can act in multiple extrapituitary tissues via autocrine/paracrine mechanisms. Protein for GnRH and mRNA for both GnRH and its receptor have been detected in human uterine endometrium and oviduct as well as in embryos at the morula/blastocyst stage in the mouse and human. Therefore, we hypothesized that GnRH may have a critical role in the development of pre-implantation embryos. To address this question, we examined the effect of a GnRH agonist and antagonist on the development of mouse embryos in vitro. For these studies, 1-cell embryos were randomly allocated to culture in KSOM containing the appropriate treatment for 144 h at 37°C in a 5% CO2 in air environment. The medium was changed every 12 h and embryos were scored daily for development. The data were compared using a χ2 test. First, we wanted to determine if a GnRH agonist, histrelin, could enhance embryonic development. Embryos were cultured with (n = 35) or without (n = 36) 10 μM histrelin. The addition of histrelin did not increase morula or blastocyst formation v. control. Second, we cultured embryos in the presence of different concentrations (0, 0.001, 0.01, 0.1, 1, and 10 μM) of the GnRH antagonist, SB-75 (cetrorelix; n = 22/treatment) in order to determine its effect on embryonic development. The 10 μM SB-75 treatment blocked embryo development beyond the compact morula stage (P < 0.001). To determine if this was a receptor mediated effect, we attempted to rescue development of SB-75 treated embryos with a histrelin challenge. Our treatments consisted of control (n = 30), 10 μM histrelin (n = 27), 10 μM SB-75 (n = 29), and 10 μM SB-75 in combination with either 1 μM (n = 27) or 10 μM (n = 25) histrelin. Both levels of histrelin partially rescued the inhibition of blastocyst formation by SB-75 (P < 0.01). Next, we were interested in examining the signaling cascade activated following binding of GnRH to its receptor in pre-implantation embryos. Toward this end, we treated embryos with inhibitors of either PKC or PKA. First, embryos were cultured in the presence of 0 (n = 33), 0.1 (n = 35), 1 (n = 35), or 10 (n = 35) μM GF109203X (GFX), a PKC inhibitor. Similar to the results obtained with SB-75, treatment with 10 μM GFX significantly reduced development to the compact morula stage and completely blocked blastocyst formation. Second, we treated embryos (n = 15 to 17/treatment) with different concentrations (0, 0.01, 0.1, 0.5, or 1 mM) of the PKA inhibitor, SQ22536. In contrast to treatment with GFX, rates of blastocyst formation were decreased only by 35% (P < 0.05) at the highest concentration of SQ22536. The percentage of embryos developing to the hatched blastocyst stage was decreased in a dose-dependent manner following SQ22536 treatment (P < 0.05); however, this effect was not consistent with SB-75 inhibition of blastocyst formation. We suggest that GnRH has an important autocrine effect on early embryonic development, potentially signaling via PKC. Funding for M Montagner was provided by CAPES, Brazil.

scholarly journals TINGKAT PERKEMBANGAN AWAL EMBRIO SAPI IN VITRO MENGGUNAKAN MEDIA TUNGGAL BERBAHAN DASAR TISSUE CULTURE MEDIUM (TCM) 199

2013 ◽  
Vol 7 (2) ◽  
Author(s):  
Mohamad Agus Setiadi ◽  
Ni Wayan Kurniani Karja
Keyword(s):  
Tissue Culture ◽  
Bovine Serum Albumin ◽  
Chorionic Gonadotropin ◽  
Bovine Serum ◽  
Culture Medium ◽  
Fetal Bovine Serum ◽  
Tissue Culture Medium ◽  
Serum Gonadotropin ◽  
Fetal Bovine

Penelitian ini bertujuan mengetahui kemampuan perkembangan awal embrio sapi in vitro menggunakan media tunggal untuk maturasi, fertilisasi, dan kultur berbahan dasar tissue culture medium (TCM) 199. Oosit sapi dikumpulkan dari rumah potong hewan dengan teknik aspirasi dan diklasifikasikan berdasarkan kekompakan sel kumulus dan sitoplasma yang homogen. Oosit dimaturasi pada medium TCM 199 yang disuplementasi dengan 10 IU/ml pregnant mare’s serum gonadotropin (PMSG), 10 IU/ml human chorionic gonadotropin (hCG), dan 10% fetal bovine serum (FBS), dilakukan selama 24 jam pada inkubator 5% CO2, 39 C. Fertilisasi dilakukan pada dua media yang berbeda yaitu media rutin fertilisasi dan media berbahan dasar TCM 199 dengan suplemen bovine serum albumin (BSA) dan heparin. Setelah fertilisasi, kumulus sel dihilangkan (denudasi), kemudian dikultur pada media TCM 199 yang disuplementasi dengan asam amino esensial dan non-esensial serta 10% FBS selama 3 hari. Hasil penelitian menunjukkan tingkat maturasi oosit pada sistem yang digunakan mampu mendukung 81,5% oosit mencapai tahap metafase II (M-II). Tingkat pembelahan embrio lebih tinggi pada media rutin dibandingkan dengan media TCM 199 yakni masing-masing 44,4 dan 23,2%. Jumlah embrio tahap 4-8 sel pada kedua perlakuan tidak berbeda nyata. Dapat disimpulkan media tunggal berbasis TCM dapat digunakan untuk produksi embrio in vitro.

Attempts at in vitro fertilization and culture of in vitro matured oocytes in sei (Balaenoptera borealis) and Bryde's (B. edeni) whales

Zygote ◽  
2009 ◽  
Vol 17 (1) ◽  
pp. 19-28 ◽  
Author(s):  
M. M. U. Bhuiyan ◽  
Y. Suzuki ◽  
H. Watanabe ◽  
K. Matsuoka ◽  
Y. Fujise ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Bovine Serum Albumin ◽  
Embryo Culture ◽  
Culture Medium ◽  
Cumulus Cells ◽  
Protein Supplementation ◽  
Vitro Fertilization ◽  
Embryo Culture Medium ◽  
Morula Stage

SummaryThe cumulus–oocyte–complexes (COCs) recovery rates with respect to reproductive status per sei (Balaenoptera borealis) and Bryde's (B. edeni) whales were determined in Experiment 1. The number of COCs recovered ranged from 16.0 to 30.6 and from 6.7 to 26.8 per sei and Bryde's whales, respectively. The effects of COCs grades and protein supplementation in embryo culture medium on development of in vitro fertilized (IVF) embryos were evaluated in sei and Bryde's whales in Experiment 2. The COCs were classified into either Grade A (COCs with five or more layers of compact cumulus cells) or Grade B (COCs with less than five layers of compact or expanded cumulus cells) before being cultured for IVM. The cleavage (12.0 to 19.5%), 4-cell (8.0 to 12.0%) and 8-cell (4.0 to 8.0%) formation rates in sei whales did not vary significantly between embryos derived from either grade A or B oocytes and between embryos cultured in either fetal whale serum (FWS)- or bovine serum albumin (BSA)-supplemented medium. The cleavage (4.0 to 14.8%), 4-cell (0.0 to 7.5%) and 8-cell (0.0 to 2.6%) formation rates in Bryde's whales did not vary significantly between embryos derived from either grade A or B oocytes and between embryos cultured in either FWS- or BSA-supplemented medium. The grade B oocytes cultured in FWS-supplemented medium developed to morula stage (1.1%) in sei whales. In conclusion, the present study indicates that IVF in sei whales is possible to achieve cleaved embryos developing to morula stage. This is the first in vitro embryo production attempt in sei and Bryde's whales.

A Precipitin-like Reaction of the Hemolymph of the Lobster Homarus americanus

1969 ◽  
Vol 26 (5) ◽  
pp. 1392-1397 ◽  
Author(s):  
James E. Stewart ◽  
Diane M. Foley
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Serum Proteins ◽  
Homarus Americanus ◽  
Test Period ◽  
Fluorescent Material

The levels of fluorescent material in the hemolymph of lobsters injected with serum proteins from lobster hemolymph labelled with fluorescein remained relatively constant over a 6-day test period; the levels in lobsters injected with bovine serum albumin labelled with fluorescein declined rapidly. A precipitin-like reaction was observed when lobster hemolymph serum was titrated with bovine serum albumin in vitro.

Preparation and in vitro photodynamic activities of novel axially substituted silicon (IV) phthalocyanines and their bovine serum albumin conjugates

2006 ◽  
Vol 16 (9) ◽  
pp. 2450-2453 ◽  
Author(s):  
Xiong-Jie Jiang ◽  
Jian-Dong Huang ◽  
Yu-Jiao Zhu ◽  
Fen-Xiang Tang ◽  
Dennis K.P. Ng ◽  
...  
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum

scholarly journals N-Acetylcysteine and α-cyano-4-hydroxycinnamic acid alter protein kinase C (PKC)-δ and PKC-ζ and diminish dysmorphogenesis in rat embryos cultured with high glucose in vitro

2007 ◽  
Vol 192 (1) ◽  
pp. 207-214 ◽  
Author(s):  
Mattias Gäreskog ◽  
Parri Wentzel
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
High Glucose ◽  
Culture Medium ◽  
Hydroxycinnamic Acid ◽  
Oxygen Species ◽  
Rat Embryos ◽  
Protein Kinase C Inhibitors ◽  
Kinase C

Malformations and growth disturbances are two- to threefold more common in infants of diabetic mothers than in offspring of non-diabetic pregnancy. Several suggestions have emerged to explain the reasons for diabetic embryopathy, including enhanced mitochondrial production of reactive oxygen species leading to altered activation of protein kinase C. This study aimed to evaluate the effect of α-cyano-4-hydroxycinnamic acid (CHC) and N-acetylcysteine (NAC) addition on morphology and activity of protein kinase C-δ and protein kinase C-ζ in rat embryos exposed to a high glucose concentration in vitro. Day 9 embryos from normal rats were cultured in 10 or 30 mM glucose concentrations with or without supplementation of CHC, NAC, or protein kinase C inhibitors specific for protein kinase C-δ and protein kinase C-ζ. Embryos were evaluated for malformations, crown rump length, and somite number. Protein kinase C-δ and protein kinase C-ζ activities were estimated by western blot by separating membranous and cytosolic fractions of the embryo. We found increased malformations and growth retardation in embryos cultured in high versus low glucose concentrations. These abnormalities were diminished when CHC and NAC or specific protein kinase C-inhibitors were added to the culture medium. The activities of embryonic protein kinase C-δ and protein kinase C-ζ were increased in the high glucose environment after 24-h culture, but were normalized by the addition of CHC and NAC as well as respective inhibitor to the culture medium. These findings suggest that mitochondrial overproduction of reactive oxygen species is involved in diabetic embryopathy. Furthermore, such overproduction may affect embryonic development, at least partly, by enhancing the activities of protein kinase C-δ and protein kinase C-ζ.

Bovine serum albumin contained in culture medium used in artificial insemination is an important anaphylaxis risk factor

2008 ◽  
Vol 90 (5) ◽  
pp. 2013.e17-2013.e19 ◽  
Author(s):  
Juan A. Pagán ◽  
Idoia Postigo ◽  
Jorge R. Rodríguez-Pacheco ◽  
Maribel Peña ◽  
Jorge A. Guisantes ◽  
...  
Keyword(s):  
Risk Factor ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Artificial Insemination ◽  
Bovine Serum ◽  
Culture Medium

Effect of yolk-sac antibody on rat embryos grown in culture

Development ◽  
1972 ◽  
Vol 27 (3) ◽  
pp. 543-553
Author(s):  
D. A. T. New ◽  
R. L. Brent
Keyword(s):  
Direct Contact ◽  
Culture Medium ◽  
Gamma Globulin ◽  
Yolk Sac ◽  
Amniotic Cavity ◽  
Pregnant Rats ◽  
Rat Embryos ◽  
Visceral Yolk Sac ◽  
Primary Action

Rat embryos, explanted with their embryonic membranes during the early stages of organogenesis ( days gestation), were grown in culture in roller tubes. Yolk-sac antibody (sheep anti rat yolk-sac gamma globulin), known to be teratogenic when injected into pregnant rats, was added to the culture medium. At concentrations of 0·1 mg/ml or more the antibody caused gross retardation of growth and differentiation. Injection of antibody into the amniotic cavity so that it had direct contact with the embryo, or between the amnion and yolk sac so that it was in contact with the mesodermal surface of the yolk sac, had little or no effect on development of the embryo or its membranes. These in vitro experiments indicate that yolk-sac antibody has an effect on development independent of any immunological reaction of the mother, and the primary action is probably on the visceral yolk-sac endoderm.

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