Effect of culture in vitro and organ culture on the dry mass of preimplantation mouse embryos

1994 ◽  
Vol 6 (2) ◽  
pp. 229 ◽  
Author(s):  
K Turner ◽  
AW Rogers ◽  
EA Lenton
Keyword(s):  
Organ Culture ◽  
Culture System ◽  
Dry Mass ◽  
Mouse Embryos ◽  
Preimplantation Embryos ◽  
Cell Stage ◽  
Organ Culture System ◽  
Preimplantation Mouse

The dry mass of mouse embryos cultured in vitro in medium alone or in an organ culture system were measured by means of the Vickers M86 scanning microinterferometer. The data were compared with previous data on the dry mass of preimplantation embryos in vivo. The metabolism of embryos cultured in vitro differs from that of fresh embryos. In cultured embryos, dry mass decreases throughout the 2-cell stage whereas the dry mass is increasing at this stage in vivo. Embryos in an organ culture system regain a dry mass profile, similar to that observed in vivo at the late cleavage stage. These results support the view that conditions for embryo metabolism are suboptimal in vitro and that, although the oviduct may confer some advantage on developing embryos in vitro, it is unable fully to support the pattern of metabolism, as assessed by dry mass, observed in vivo.

In vivo and in vitro Study of the Effects of Vitamin A Deficiency on Rat Third Molar Development

1986 ◽  
Vol 65 (12) ◽  
pp. 1445-1448 ◽  
Author(s):  
S.S. Harris ◽  
J.M. Navia
Keyword(s):  
Vitamin A ◽  
Organ Culture ◽  
Culture System ◽  
Vitamin A Deficiency ◽  
Third Molar ◽  
In Vitro Study ◽  
Organ Culture System ◽  
Vitamin A Status

We have examined the effect of in vivo vitamin A status on subsequent rat third molar formation and mineralization in an in vitro organ culture system. Vitamin A deficiency imposed during an eight-day in vitro period caused effects very similar to those of vitamin A deficiency imposed on rats in vivo. Analysis of the data also demonstrates that retinoic acid is capable of reversing the interference in mineralization of third molars induced by vitamin A deficiency in the organ culture system.

Development of Sensory Structures in Organ Cultures of the Twelfth and Thirteenth Gestation Day Mouse Embryo Inner Ears

1973 ◽  
Vol 82 (4_suppl) ◽  
pp. 3-18
Keyword(s):  
Organ Culture ◽  
Mouse Embryo ◽  
Culture System ◽  
Organ Of Corti ◽  
Sensory Cells ◽  
Equal Distribution ◽  
Organ Culture System ◽  
Sensory Structures

Twelfth and thirteenth gestation day mouse embryo otocysts have been explanted into an organ culture system that promotes advances in morphogenesis and differentiation of sensory structures. The pattern of morphogenesis that occurs “in vitro” is not equivalent to that which occurs in the “in vivo” environment. These morphogenetic changes occur with greatest frequency in the explanted thirteenth gestation day otocyst. The development of sensory structures occurs with equal distribution in the twelfth and thirteenth gestation day explanted otocysts. The thirteenth gestation day mouse otocyst favors the development of organ of Corti type formations, and the twelfth gestation day otocyst favors the development of maculae of sensory cells of a vestibular character in the organ culture system employed. The thirteenth gestation day otocyst requires a shorter period of “in vitro” development to produce differentiation of sensory structures. The sensory structures that develop “in vitro” follow the pattern of the sensory structures that develop “in vivo.”

scholarly journals Ontogenic development of B-lymphocyte function and tolerance susceptibility in vivo and in an in vitro organ culture system.

1980 ◽  
Vol 151 (2) ◽  
pp. 429-445 ◽  
Author(s):  
J M Teale ◽  
T E Mandel
Keyword(s):  
B Cells ◽  
Organ Culture ◽  
Culture System ◽  
B Lymphocyte ◽  
Lymphocyte Function ◽  
Organ Culture System ◽  
Ontogenic Development ◽  
In Vitro Organ Culture

The maturation of B-lymphocyte function during fetal development was studied in vivo and in an in vitro organ culture system. The results indicated that the progenitors for 2,4-dinitrophenol (DNP)-specific B cells are present as early as 14 d of gestation in liver and possibly as early as 15 d in spleen. In addition, it was found that the organ culture system supports the development of B lymphocytes as measured by an increase in both the percentage of surface immunoglobulin-positive cells and the frequency of clonable DNP-specific B cells after culturing. The majority of anti-DNP-secreting clones resulting from the antigenic stimulation of fetal B cells produced only the IgM isotype, and the ability to secrete the IgG isotypes increased as a function of gestational age. Because fetal DNP precursors from spleens and livers that had been incubated in organ culture resulted in a greater proportion of clones secreting IgG compared with age-matched uncultured controls, it was concluded that the maturation with regard to the ability to secrete IgG can occur in vitro. In studies relating to the ontogenetic development of tolerance susceptibility, it was found that up to one-half of the DNP-specific B-cell precursors from livers and spleens less than 18 or 19 d of gestation were resistant to tolerogen treatment for 24 h as if in a pretolerant phase. However, if tolerogen were present for 3--5 d during organ culture there was near total elimination of potential DNP clones. This finding suggested that the 24-h induction period was insufficient for affecting the DNP-specific precursors in livers and spleens from the earlier gestational ages, and that a proportion of precursors could subsequently form DNP clones in the splenic focus assay after the removal of tolerogen.

Development of Sensory Structures in Organ Cultures of the Twelfth and Thirteenth Gestation Day Mouse Embryo Inner Ears

1973 ◽  
Vol 82 (1) ◽  
pp. 1-18 ◽  
Author(s):  
Thomas R. Van De Water ◽  
P. Heywood ◽  
Robert J. Ruben
Keyword(s):  
Organ Culture ◽  
Mouse Embryo ◽  
Culture System ◽  
Organ Of Corti ◽  
Sensory Cells ◽  
Equal Distribution ◽  
Organ Culture System ◽  
Sensory Structures

Twelfth and thirteenth gestation day mouse embryo otocysts have been explanted into an organ culture system that promotes advances in morphogenesis and differentiation of sensory structures. The pattern of morphogenesis that occurs “in vitro” is not equivalent to that which occurs in the “in vivo” environment. These morphogenetic changes occur with greatest frequency in the explanted thirteenth gestation day otocyst. The development of sensory structures occurs with equal distribution in the twelfth and thirteenth gestation day explanted otocysts. The thirteenth gestation day mouse otocyst favors the development of organ of Corti type formations, and the twelfth gestation day otocyst favors the development of maculae of sensory cells of a vestibular character in the organ culture system employed. The thirteenth gestation day otocyst requires a shorter period of “in vitro” development to produce differentiation of sensory structures. The sensory structures that develop “in vitro” follow the pattern of the sensory structures that develop “in vivo.”

DNA synthesis in the preimplantation mouse embryo

Development ◽  
1972 ◽  
Vol 27 (2) ◽  
pp. 431-445
Author(s):  
P. Barlow ◽  
D. A. J. Owen ◽  
Chris Graham
Keyword(s):  
S Phase ◽  
Preimplantation Embryos ◽  
Maternal Environment ◽  
Cell Stage ◽  
Cell Cycles ◽  
Preimplantation Mouse Embryo ◽  
Preimplantation Mouse ◽  
Dna Amounts

Strain PO preimplantation embryos were labelled with [3H]thymidine. The incorporation of the label was studied by autoradiography of air-dried and serially sectioned embryos. DNA amounts were measured with a microdensitometer. The following observations were made at the 5- to 16-cell stages. All nuclei contained 2C–4C amounts of DNA and all could eventually synthesize DNA. However, after short labelling intervals, unlabelled nuclei were found with 2C and AC amounts of DNA. We concluded that both the G1 and the G2 phases of the cell cycle were present at this time. Embryos were found in which the S phase of the 4th and the 5th cell cycles post fertilization overlapped. In 12- to 15-cell embryos which contained inside cells it was found that the inside cells were produced by one of the first four 8-cell-stage blastomeres to divide. The inside cells of 9- to 256-cell embryos had a significantly higher labelling index than the outside cells and the number of inside cells increased faster than the number of outside cells during development. We concluded that the inside cells were dividing faster than the outside cells. Blastocysts which had developed in vivo or in vitro contained nuclei with greater than 4C amounts of DNA. We concluded that the development of excess DNA amounts does not depend on the maternal environment. These nuclei which contained greater than 4C amounts of DNA were labelled after short exposures to radioactivity. We concluded that it was likely that they were becoming polyploid.

scholarly journals Effects of embryo culture on global pattern of gene expression in preimplantation mouse embryos

Reproduction ◽  
2004 ◽  
Vol 128 (3) ◽  
pp. 301-311 ◽  
Author(s):  
Paolo Rinaudo ◽  
Richard M Schultz
Keyword(s):  
Gene Expression ◽  
Preimplantation Embryos ◽  
Expression Analysis Systematic Explorer ◽  
Cell Stage ◽  
Global Pattern ◽  
Preimplantation Mouse Embryos ◽  
Preimplantation Mouse ◽  
Global Patterns ◽  
The One

Culture of preimplantation embryos affects gene expression. The magnitude of the effect on the global pattern of gene expression, however, is not known. We compared global patterns of gene expression in blastocysts cultured from the one-cell stage in either Whitten’s medium or KSOM + amino acids (KSOM/AA) with that of blastocysts that developed in vivo, using the Affymetrix MOE430A chip. The analysis revealed that expression of 114 genes was affected after culture in Whitten’s medium, whereas only 29 genes were mis-expressed after culture in KSOM/AA. Expression Analysis Systematic Explorer was used to identify biological and molecular processes that are perturbed after culture and indicated that genes involved in protein synthesis, cell proliferation and transporter function were down-regulated after culture in Whitten’s medium. A common set of genes involved in transporter function was also down-regulated after culture in KSOM/AA. These results provide insights as to why embryos develop better in KSOM/AA than in Whitten’s medium, and highlight the power of microarray analysis to assess global patterns of gene expression.

Localization of cytoskeletal proteins in preimplantation mouse embryos

Development ◽  
1980 ◽  
Vol 55 (1) ◽  
pp. 211-225
Author(s):  
E. Lehtonen ◽  
R. A. Badley
Keyword(s):  
Blastocyst Stage ◽  
Cytoskeletal Proteins ◽  
Mouse Embryos ◽  
Trophectoderm Cells ◽  
Apparent Concentration ◽  
Preimplantation Mouse ◽  
Early Mouse ◽  
Filament Protein

The immunofluorescence technique was used to detect the presence and distribution of actin, alpha-actinin, tubulin and 10 nm filament protein in early mouse embryos. Actin and alpha-actinin stainings showed a distinct concentration to a peripheral layer in the cleavage-stage blastomeres and in trophectoderm cells. Dots of fluorescence appeared in this cortical staining pattern. The distribution of tubulin staining in the blastomere cytoplasm was relatively even with apparent concentration at the perinuclear region and frequently at wide intercellular contact areas. 10 nm filament protein was distributed evenly in the blastomere cytoplasm without cortical concentration of the label. At the blastocyst stage, the trophectoderm cells in blastocyst outgrowths in vitro developed well organized cytoskeletons including both microfilament, microtubule and 10 nm filament elements. Comparable structures were not observed in blastocysts in vivo, or in late hatched blastocysts cultured in suspension. The morphogenetic significance of the observations is discussed.

180 IDENTIFICATION AND QUANTIFICATION OF DIFFERENTIALLY REPRESENTED TRANSCRIPTS IN PREIMPLANTATION BOVINE EMBRYOS

2008 ◽  
Vol 20 (1) ◽  
pp. 169 ◽  
Author(s):  
C. E. McHughes ◽  
G. K. Springer ◽  
L. D. Spate ◽  
R. Li ◽  
R. J. Woods ◽  
...  
Keyword(s):  
Relative Abundance ◽  
Nuclear Transfer ◽  
Developmental Stages ◽  
Reproductive Technologies ◽  
Blastocyst Stage ◽  
Preimplantation Embryos ◽  
Cell Stage ◽  
Production Techniques

Identification of transcripts that are present at key development stages of preimplantation embryos is critical for a better understanding of early embryogenesis. To that end, this project had two goals. The first was to characterize the relative abundance of multiple transcripts during several developmental stages, including metaphase II-stage oocytes (MPII), and 2-cell-stage (2-cell), precompact morula (PCM), and in vitro-produced blastocyst-stage (IVTBL) embryos. The second was to characterize differences in the relative abundance of transcripts present in in vivo- (IVVBL), in vitro-, and nuclear transfer-produced (NTBL) blastocysts. It was our hypothesis that the identification of differentially represented transcripts from these stages would reveal not only developmentally important genes, but also genes that might be aberrantly expressed due to embryo production techniques. Individual clusters from a large bovine EST project (http://genome.rnet.missouri.edu/Bovine/), which focused on female reproductive tissues and embryos, were compared using Fisher's exact test weighted by number of transcripts per tissue by gene (SAS PROC FREQ; SAS Institute, Inc., Cary, NC, USA). Of the 3144 transcripts that were present during embryogenesis, 125 were found to be differentially represented (P < 0.01) in at least one pairwise comparison (Table 1). Some transcripts found to increase in representation from the MPII to the 2-cell stage include protein kinases, PRKACA and CKS1, as well as the metabolism-related gene, PTTG1. These same transcripts were also found to decrease in representation from the 2-cell to the PCM stage. RPL15 (translation) and FTH1 (immune function) were both more highly represented in the PCM than in the 2-cell stage. From PCM to IVTBL, we saw an increase in RPS11, another translation-related transcript. When comparing blastocyst-stage embryos from different production techniques, several transcripts involved in energy production (e.g., COX7B and COX8A) were found to be more highly represented in the NTBL than in the IVTBL. COX8A was also more highly represented in the IVVBL than in the IVTBL. By investigating these differentially represented transcripts, we will be able to better understand the developmental implications of embryo manipulation. We may also be able to better develop reproductive technologies that lead to in vitro- and nuclear transfer-derived embryos which more closely follow a normal program of development. Table 1. Differentially represented transcripts between developmental stages

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