scholarly journals Identification of embryonic cell lineages in histological sections of M. musculus ↔ M. caroli chimaeras

Development ◽  
1983 ◽  
Vol 73 (1) ◽  
pp. 179-191
Author(s):  
J. Rossant ◽  
M. Vijh ◽  
L. D. Siracusa ◽  
V. M. Chapman
Keyword(s):  
Inner Cell Mass ◽  
Cell Mass ◽  
Embryonic Cell ◽  
Dna Probe ◽  
Marker System ◽  
Cell Lineages ◽  
Mus Caroli ◽  
First Time ◽  
Derivatives Of

An in situ cell marker system has been developed which allows identification of Mus caroli and Mus musculus cells in interspecific chimaeras. A radioactively labelled, cloned DNA probe to M. musculus satellite DNA was hybridized in situ to sections of M. musculus and M. caroli adult tissues. Autoradiography revealed high levels of hybridization to the nuclei of M. musculus cells, but little or no label bound to M. caroli cells. The DNA probe could also distinguish M. musculus and M. caroli cells in the same tissue section. Patches of labelled and unlabelled cells were clearly identified in sections of adult chimaeric tissues and also in the embryonic ectoderm of 6·5-day embryonic chimaeras. The ability to recognize M. musculus and M. caroli cells in sections of chimaeras should provide a powerful new tool in analyses of cell lineages in both embryonic and adult mouse chimaeras. The marker system has several advantages over other marker systems so far developed, the most important of which is its ubiquity. Since it is a nuclear marker, only cells without nuclei should be unsuited to its use. The potential of the marker system has been shown by its use in demonstrating directly for the first time the postimplantation derivatives of inner cell mass and trophectoderm in blastocysts ‘reconstituted’ with M. musculus trophectoderm and M. caroli inner cell mass.

Interspecific cell markers and lineage in mammals

1985 ◽  
Vol 312 (1153) ◽  
pp. 91-100 ◽  
Keyword(s):  
Dna Hybridization ◽  
Cell Lineage ◽  
Embryonic Cell ◽  
Mammalian Embryo ◽  
Marker System ◽  
Cell Lineages ◽  
Streptavidin Peroxidase ◽  
Derivatives Of ◽  
Lineage Analysis

Study of cell lineage in the mammalian embryo has relied heavily on the use of chimeras to follow the fate of genetically marked cells in later development. Such studies have often been limited by the types of genetic markers available; there are very few markers that allow analysis of the spatial distribution of individual cells at all stages of development. We have developed a marker system that is based on the identification of cells of Mus musculus origin in M. musculus-M. caroli chimeras by in situ DNA-DNA hybridization using a cloned probe to M. musculus satellite DNA. This provides the first ubiquitous in situ cell marker system for mammalian chimeras. We have recently refined the system by the use of biotin-labelled probes and detection of hybridization by streptavidin-peroxidase binding. This increases both the speed and the resolution of the assay. We have used the marker for cell lineage analysis in both embryonic and adult chimeras and results from analysis of the derivatives of early cell lineages in later development and study of coherent growth versus cell mixing in the postimplantation embryo are presented. The importance of understanding embryonic cell lineages as a prelude to molecular studies is emphasized.

Lineage analysis of inner cell mass and trophectoderm using microsurgically reconstituted mouse blastocysts

Development ◽  
1982 ◽  
Vol 68 (1) ◽  
pp. 199-209
Author(s):  
Virginia E. Papaioannou
Keyword(s):  
Enzyme Assay ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Glucose Phosphate Isomerase ◽  
Mouse Blastocyst ◽  
Cell Lineages ◽  
Glucose Phosphate ◽  
Parietal Endoderm ◽  
First Time ◽  
Lineage Analysis

The fate of mouse blastocyst tissues was examined following reconstitution of blastocystsfrom isolated inner cell mass (ICM) and trophectoderm differing for electrophoretic variantsat the glucose phosphate isomerase (GPI-1) locus. A modified microsurgical method was usedand a more sensitive enzyme assay allowed finer dissection of developing chimaeric con-ceptuses. In seven of nine cases, the extraembryonic ectoderm or the later ectoderm of thechorion was entirely of the blastocyst trophectoderm enzyme type, providing the first ditectevidence that this tissue can be wholly derived from the trophectoderm. The two exceptionscould represent contamination of the ICM with trophectoderm or might indicate somedevelopmental lability of ICM cells. In addition, the results confirm the cell lineages of othertissues of the 7·5- to 9·5-day pc embryo and, for the first time, directly demonstrate the ICMorigin of the parietal endoderm.

Trophectodermal processes regulate the expression of totipotency within the inner cell mass of the mouse expanding blastocyst

Development ◽  
1984 ◽  
Vol 84 (1) ◽  
pp. 63-90
Author(s):  
Tom P. Fleming ◽  
Paul D. Warren ◽  
Julia C. Chisholm ◽  
Martin H. Johnson
Keyword(s):  
Inner Cell Mass ◽  
Cell Mass ◽  
Central Area ◽  
Ultrastructural Changes ◽  
Trophectoderm Cells ◽  
Junctional Complexes ◽  
Secondary Lysosomes ◽  
Cytoplasmic Processes ◽  
Cellular Cover

Mouse blastocysts, aged 0, 2, 6 and 12 h from the onset of cavitation, were examined by transmission (TEM) and scanning (SEM) electron microscopy. In TEM sections, trophectoderm cells (TE) differed morphologically from those of the inner cell mass (ICM) by their flattened shape, paler cytosol staining and polarized disposition of both junctional complexes (apicolateral) and intracellular secondary lysosomes (SL; basal). Throughout this period of development, cytoplasmic processes, characterized by abundant SLs, cover approximately 80 % of the juxtacoelic face of the ICM. These processes are shown to be derived from the basal surface of TE cells intermediately placed between polar and mural regions. In SEM preparations of the juxtacoelic ICM surface, revealed by ‘cracking open’ blastocysts, the processes appear as tongue-shaped, centripetally oriented structures which terminate collectively at a central area on the ICM surface. The potential of cultured ICMs to generate TE was demonstrated following their immunosurgical isolation from blastocysts aged up to 12 h post cavitation and by examining the sequence of ultrastructural changes associated with TE generation by ICMs from 2 h blastocysts. In contrast, the juxtacoelic cells of similarly aged ICMs observed in situ in ultrasections of intact embryos showed little or no evidence of totipotency expression as judged by the absence of TE characteristics. Since TE expression within presumptive ICM cells is thought to be generated by an asymmetry of cell contacts (Johnson & Ziomek, 1983), we propose that the juxtacoelic TE processes, by providing a cellular cover to the ICM, function in suppressing the expression in situ of ICM totipotency.

Stem cell defects in parthenogenetic peri-implantation embryos

Development ◽  
1995 ◽  
Vol 121 (7) ◽  
pp. 2069-2077
Author(s):  
E.D. Newman-Smith ◽  
Z. Werb
Keyword(s):  
Stem Cells ◽  
Growth Factor ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Cell Types ◽  
Insulin Like Growth Factor ◽  
Embryonic Antigen ◽  
Parietal Endoderm

Mouse embryos containing only maternal chromosomes (parthenotes) develop abnormally in vivo, usually failing at the peri-implantation stage. We have analyzed the development of parthenote embryos by using an inner cell mass (ICM) outgrowth assay that mimics peri-implantation development. ICMs from normal embryos maintained undifferentiated stem cells positive for stage-specific embryonic antigen-1 and Rex-1 while differentiating into a variety of cell types, including visceral endoderm-like cells and parietal endoderm cells. In contrast, ICMs from parthenotes failed to maintain undifferentiated stem cells and differentiated almost exclusively into parietal endoderm. This suggests that parthenote ICMs have a defect that leads to differentiation, rather than maintenance, of the stem cells, and a defect that leads to a parietal endoderm fate for the stem cells. To test the hypothesis that the ICM population is not maintained owing to a lack of proliferation of the stem cells, we investigated whether mitogenic agents were able to maintain the ICM population in parthenotes. When parthenote blastocysts were supplied with the insulin-like growth factor-1 receptor (Igf-1r) and insulin-like growth factor-2 (Igf-2), two genes not detectable in parthenote blastocysts by in situ hybridization, the ICM population was maintained. Similarly, culture of parthenote blastocysts in medium conditioned by embryonic fibroblasts and supplemented with the maternal factor leukemia inhibitory factor maintained the ICM population. However, once this growth factor-rich medium was removed, the parthenote ICM cells still differentiated predominantly into parietal endoderm.(ABSTRACT TRUNCATED AT 250 WORDS)

Isolation of the murine cyclin B2 cDNA and characterization of the lineage and temporal specificity of expression of the B1 and B2 cyclins during oogenesis, spermatogenesis and early embryogenesis

Development ◽  
1993 ◽  
Vol 118 (1) ◽  
pp. 229-240 ◽  
Author(s):  
D.L. Chapman ◽  
D.J. Wolgemuth
Keyword(s):  
Germ Cell ◽  
Germ Cells ◽  
Northern Blot ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Cell Development ◽  
Early Embryogenesis ◽  
Northern Blot Hybridization ◽  
Germ Cell Development

A cDNA encoding the murine cyclin B2 (cycB2) was isolated from an adult mouse testis cDNA library as part of studies designed to identify cyclins involved in murine germ cell development. This cycB2 cDNA was then used to examine the pattern of cycB2 expression during male and female germ cell development and in early embryogenesis, and to compare this expression with the previously characterized expression of cycB1. A single 1.7 kb cycB2 transcript was detected by northern blot hybridization analysis of total RNA isolated from midgestation embryos and various adult tissues. Northern blot and in situ hybridization analyses revealed that cycB2 expression in the testis was most abundant in the germ cells, specifically in pachytene spermatocytes. This is in contrast to the highest levels of expression of cycB1 being present in early spermatids. In situ analysis of the ovary revealed cycB2 transcripts in both germ cells and somatic cells, specifically in the oocytes and granulosa cells of growing and mature follicles. The pattern of cycB1 and cycB2 expression in ovulated and fertilized eggs was also examined. While the steady state level of cycB1 and cycB2 signal remained constant in oocytes and ovulated eggs, signal of both appeared to decrease following fertilization. In addition, both cycB1 and cycB2 transcripts were detected in the cells of the inner cell mass and the trophectoderm of the blastocyst. These results demonstrate that there are lineage- and developmental-specific differences in the pattern of the B cyclins in mammalian germ cells, in contrast to the co-expression of B cyclins in the early conceptus.

scholarly journals Cytoplasmic String Between ICM and mTE Is a Positive Predictor of Clinical Pregnancy and Live Birth Outcomes in Elective Frozen-thawed Single Blastocyst Transfer Cycles: a Time-lapse Study

2020 ◽  
Author(s):  
Bing-Xin Ma ◽  
Lei Jin ◽  
Bo Huang
Keyword(s):  
Birth Outcomes ◽  
Live Birth ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Time Lapse ◽  
Clinical Pregnancy ◽  
Blastocyst Transfer ◽  
Single Blastocyst Transfer ◽  
First Time ◽  
Present Group

Abstract Background: In this study, we aim to investigate whether cytoplasmic string between inner cell mass (ICM) and mural trophectoderm (mTE) is a positive predictor of clinical pregnancy and live birth outcomes.Methods: 1,267 elective frozen-thawed single blastocyst transfer (eSBT) cycles cultured in time-lapse incubation system from January 2018 to May 2019 were involved in the study. Blastocysts were grouped according to the appearance of cytoplasmic strings between ICM and mTE cells, and identified as “Present” and “Absent” groups. In Present group, they were further categorized according to the quantity of cytoplasmic strings between ICM and mTE cells. Clinical pregnancy and live birth outcomes of blastocysts were used to evaluate the effect of cytoplasmic strings between ICM and mTE.Results: The baseline demographic and laboratory features were similar between the Present and Absent groups of cytoplasmic strings between ICM and mTE (P>0.05). According to the time-lapse analysis, cytoplasmic strings between ICM and mTE were more visible among good quality blastocysts. Furthermore, blastocysts with cytoplasmic strings showed a higher clinical pregnancy and live birth rates (P<0.05), and no significant differences were observed in abortion rate and birth weight (P>0.05).Conclusions: Although the previous conclusions of cytoplasmic strings were controversial, the present time-lapse analysis provides the evidence for the first time that cytoplasmic strings between ICM and mTE cells would be a positive predictor of clinical pregnancy and live birth outcomes in elective frozen-thawed single blastocyst transfer cycles.

scholarly journals Cytoplasmic String Between ICM and mTE Is a Positive Predictor of Clinical Pregnancy and Live Birth Outcomes in Elective Frozen-thawed Single Blastocyst Transfer Cycles: A Time-lapse Study

2021 ◽  
Author(s):  
Bingxin Ma ◽  
Lei Jin ◽  
Bo Huang
Keyword(s):  
Birth Outcomes ◽  
Live Birth ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Time Lapse ◽  
Clinical Pregnancy ◽  
Blastocyst Transfer ◽  
Single Blastocyst Transfer ◽  
First Time ◽  
Present Group

Abstract Background: In this study, we aim to investigate whether cytoplasmic string between inner cell mass (ICM) and mural trophectoderm (mTE) is a positive predictor of clinical pregnancy and live birth outcomes.Methods: 1,267 elective frozen-thawed single blastocyst transfer (eSBT) cycles cultured in time-lapse incubation system from January 2018 to May 2019 were involved in the study. Blastocysts were grouped according to the appearance of cytoplasmic strings between ICM and mTE cells, and identified as “Present” and “Absent” groups. In Present group, they were further categorized according to the quantity of cytoplasmic strings between ICM and mTE cells. Clinical pregnancy and live birth outcomes of blastocysts were used to evaluate the effect of cytoplasmic strings between ICM and mTE.Results: The baseline demographic and laboratory features were similar between the Present and Absent groups of cytoplasmic strings between ICM and mTE (P>0.05). According to the time-lapse analysis, cytoplasmic strings between ICM and mTE were more visible among good quality blastocysts. Furthermore, blastocysts with cytoplasmic strings showed a higher clinical pregnancy and live birth rates (P<0.05), and no significant differences were observed in abortion rate and birth weight (P>0.05).Conclusions: Although the previous conclusions of cytoplasmic strings were controversial, the present time-lapse analysis provides the evidence for the first time that cytoplasmic strings between ICM and mTE cells would be a positive predictor of clinical pregnancy and live birth outcomes in elective frozen-thawed single blastocyst transfer cycles.

Genetic identification of tissue of origin of cellular populations within the mouse placenta

Development ◽  
1985 ◽  
Vol 86 (1) ◽  
pp. 177-189
Author(s):  
J. Rossant ◽  
B. A. Croy
Keyword(s):  
Genetic Markers ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Complex Mixture ◽  
Placental Tissue ◽  
Genetic Identification ◽  
Blood Capillaries ◽  
Maternal Contribution ◽  
Foetal Blood

The mouse haemochorial placenta is a complex mixture of maternal cells and foetal trophectoderm and inner cell mass (ICM)-derived cells. The majority of the placental tissue is assumed to be trophoblast in origin but the exact extent and localization of the ICM and maternal contribution has not previously been determined. Using embryo transfer and reconstituted blastocyst techniques, combined with isozymal and in situ genetic markers, we have established that about 70% of the 13 to 15-day placenta is trophectoderm-derived, 30% is maternal in origin, and 4% develops from the ICM. Nearly all of the maternal contribution was confined to the spongiotrophoblast region and all of the ICM contribution was confined to the labyrinthine trophoblast region, where it formed the foetal blood capillaries and the endodermal sinuses. Using the same genetic markers, we showed that cell suspension techniques commonly used to produce ‘trophoblast’ cell preparations from placenta do not enrich for trophoblast, and, indeed, that collagenase, the preferred dissociation technique for cell viability, produced cell suspensions in which ICM and maternal cells were preferentially dissociated. No method for producing pure trophoblast populations has yet been found. Some unusually high ICM contributions to the placenta were found in reconstituted blastocyst experiments using ICMs isolated from early 3·5-day blastocysts, suggesting that these ICMs may have contributed to the trophectoderm layer of the blastocyst. These and other experiments suggest that the inner cell mass lineage may not be closed until some time after formation of the blastocyst.

The development potential of parthenogenetically derived cells in chimeric mouse embryos: implications for action of imprinted genes

Development ◽  
1988 ◽  
Vol 104 (1) ◽  
pp. 175-182 ◽  
Author(s):  
H.J. Clarke ◽  
S. Varmuza ◽  
V.R. Prideaux ◽  
J. Rossant
Keyword(s):  
Inner Cell Mass ◽  
Cell Mass ◽  
Blastocyst Stage ◽  
Abnormal Development ◽  
Imprinted Genes ◽  
Chimeric Mouse ◽  
Paternal Genome ◽  
Transgenic Mouse Line ◽  
Parthenogenetic Embryos

Parthenogenetic embryos of mice die shortly after implantation and characteristically contain poorly developed extraembryonic tissue. To investigate the basis of the abnormal development of parthenotes, we combined them with normal embryos to produce chimeras and examined the distribution of the parthenogenetically derived cells during preimplantation and early postimplantation development. The parthenogenetic embryos were derived from a transgenic mouse line bearing a large insert, which allowed these cells to be identified in histological sections using in situ hybridization. At the blastocyst stage, the parthenogenetic embryos contributed cells to the trophectoderm (TE) and inner cell mass (ICM) of chimeras. By 6.5 days, however, in almost every embryo, parthenogenetically derived cells were not detected in the extraembryonic trophoblast tissue descended from the TE. In contrast, parthenogenetically derived cells could contribute to all descendants of the ICM of 6.5-and 7.5-day chimeras, including the extraembryonic visceral and parietal endoderm. Quantitative analysis of the degree of chimerism in the embryonic ectoderm at 6.5-7.5 days indicated that parthenogenetically derived cells could contribute as extensively as normal cells. These results indicate that normal trophoblast development requires gene expression from the paternally inherited genome before 6.5 days of embryogenesis. Tissues of the ICM lineage, however, apparently can develop independently of the paternal genome at least to 7.5 days of embryogenesis. Comparison of these results with those of others suggests that the influence of imprinted genes is manifested at different times and in a variety of tissues during development.

An in situ transgenic enzyme marker for the midgestation mouse embryo and the visualization of inner cell mass clones during early organogenesis

Development ◽  
1989 ◽  
Vol 106 (1) ◽  
pp. 37-46 ◽  
Author(s):  
R.S. Beddington ◽  
J. Morgernstern ◽  
H. Land ◽  
A. Hogan
Keyword(s):  
Cell Fate ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Early Organogenesis ◽  
Transgenic Mouse Strain ◽  
Transgenic Cells ◽  
Actin Promoter ◽  
Cell Autonomy ◽  
Lac Z

In order to study the deployment of cells during gastrulation and early organogenesis, it is necessary to have an in situ cell marker which can be used to follow cell fate. To create such a marker a transgenic mouse strain, designated Tg(Act-lac Z)-1, which carries 6 copies of the Escherichia coli lac Z gene under the control of the rat beta-actin promoter, was made by pronuclear injection of DNA. Staining early postimplantation hemizygous mouse conceptuses, during gastrulation and early organogenesis, for beta-galactosidase activity shows that lac Z expression is ubiquitous and constitutive in all epiblast derivatives of the 10th day conceptus. No activity is seen in trophectoderm and primitive endoderm derivatives. Postimplantation grafts of [3H]thymidine-labelled transgenic cells establish the cell autonomy of this transgenic marker. Preliminary observations on the distribution of inner cell mass (ICM) descendant clones, identified in situ in midgestation conceptuses, confirm the pluripotency of individual ICM cells. The implications regarding patterns of cell growth in nascent fetal primordia are discussed.

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