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

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.

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Investigation of the determinative state of the mouse inner cell mass

Development ◽

10.1242/dev.33.4.979 ◽

1975 ◽

Vol 33 (4) ◽

pp. 979-990

Author(s):

J. Rossant

Keyword(s):

Inner Cell Mass ◽

Cell Mass ◽

In Utero ◽

Glucose Phosphate Isomerase ◽

Trophoblast Cells ◽

Electrophoretic Variants ◽

Glucose Phosphate

Inner cell masses (ICMs) were dissected from 3½- and 4½-day blastocysts and cultured in contact with 2½-day morulae. Blastocysts and morulae were homozygous for different electrophoretic variants of the enzyme glucose phosphate isomerase (GPI). Aggregation of ICMs and morulae was observed, and such aggregates were able to form blastocysts in vitro and morphologically normal foetuses in utero. GPI analysis of these conceptuses revealed that most were chimaeric. However, donor ICM-type isozyme was only detected in the embryonic and extra-embryonic fractions of the chimaeras and never in the trophoblastic fraction. Thus, ICM cells appear unable to form trophoblast derivatives even when exposed to ‘outside’ conditions as experienced by developing trophoblast cells. This is evidence that ICM cells, although not overtly differentiated, are determined by 3½ days.

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Origin of the ectoplacental cone and secondary giant cells in mouse blastocysts reconstituted from isolated trophoblast and inner cell mass

Development ◽

10.1242/dev.30.3.561 ◽

1973 ◽

Vol 30 (3) ◽

pp. 561-572

Author(s):

R. L. Gardner ◽

V. E. Papaioannou ◽

S. C. Barton

Keyword(s):

Cell Transformation ◽

Inner Cell Mass ◽

Cell Mass ◽

Giant Cells ◽

Glucose Phosphate Isomerase ◽

Electrophoretic Variants ◽

Decidual Tissue ◽

Glucose Phosphate ◽

Donor Type ◽

Ectoplacental Cone

1. Inner cell mass (ICM) and trophoblast tissue were isolated from 3½-day post-coitum mouse blastocysts that were homozygous for different electrophoretic variants of the enzyme glucose phosphate isomerase (GPI). Blastocysts were reconstituted from these tissues, transferred to pseudo-pregnant recipients and allowed to develop to the early somite stage. 2. The embryo plus membranes and trophoblast were dissected and typed separately for GPI. 3. Contamination of trophoblast with maternal decidual tissue was quantified. 4. The trophoblast of the implanted embryos was almost exclusively of the trophoblastdonor GPI type. The embryos plus membranes were mainly of the ICM-donor type but most also showed a substantial proportion of trophoblast-donor type. 5. It is argued that the ICM controls trophoblast proliferation by inhibiting giant cell transformation of adjacent trophoblast cells rather than through making a significant cellular contribution.

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

Development ◽

10.1242/dev.73.1.179 ◽

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.

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When and how does cell division order influence cell allocation to the inner cell mass of the mouse blastocyst?

Development ◽

10.1242/dev.100.2.325 ◽

1987 ◽

Vol 100 (2) ◽

pp. 325-332

Author(s):

C.L. Garbutt ◽

M.H. Johnson ◽

M.A. George

Keyword(s):

Cell Division ◽

Cell Population ◽

Inner Cell Mass ◽

Cell Mass ◽

Cell Lineage ◽

The Other ◽

Mouse Blastocyst ◽

Short Term ◽

Cell Stage ◽

Cell Embryo

Aggregate 8-cell embryos were constructed from four 2/8 pairs of blastomeres, one of which was marked with a short-term cell lineage marker and was also either 4 h older (derived from an early-dividing 4-cell) or 4 h younger (derived from a late-dividing 4-cell) than the other three pairs. The aggregate embryos were cultured to the 16-cell stage, at which time a second marker was used to label the outside cell population. The embryos were then disaggregated and each cell was examined to determine its labelling pattern. From this analysis, we calculated the relative contributions to the inside cell population of the 16-cell embryo of older and younger cells. Older cells were found to contribute preferentially. However, if the construction of the aggregate 8-cell embryo was delayed until each of the contributing 2/8 cell pairs had undergone intercellular flattening and then had been exposed to medium low in calcium to reverse this flattening immediately prior to aggregation, the advantage possessed by the older cells was lost. These results support the suggestion that older cells derived from early-dividing 4-cell blastomeres contribute preferentially to the inner cell mass as a result of being early-flattening cells.

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Stem cell defects in parthenogenetic peri-implantation embryos

Development ◽

10.1242/dev.121.7.2069 ◽

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)

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Investigation of cell lineage and differentiation in the extraembryonic endoderm of the mouse embryo

Development ◽

10.1242/dev.68.1.175 ◽

1982 ◽

Vol 68 (1) ◽

pp. 175-198

Author(s):

R. L. Gardner

Keyword(s):

Inner Cell Mass ◽

Cell Mass ◽

Cell Lineage ◽

Early Embryo ◽

Visceral Endoderm ◽

Primitive Endoderm ◽

Developmental Potential ◽

Extraembryonic Endoderm ◽

Parietal Endoderm ◽

Endodermal Cells

The technique of injecting genetically labelled cells into blastocysts was used in an attempt to determine whether the parietal and visceral endoderm originate from the same or different cell populations in the early embryo. When the developmental potential of 5th day primitive ectoderm and primitive endoderm cells was compared thus, only the latter were found to colonize the extraembryonic endoderm. Furthermore, single primitive endoderm cells yielded unequivocal colonization of both the parietal and the visceral endoderm in a proportion of chimaeras. However, in the majority of primitive endodermal chimaeras, donor cells were detected in the parietal endoderm only, cases of exclusively visceral colonization being rare. Visceral endoderm cells from 6th and 7th day post-implantation embryos also exhibited a striking tendency to contribute exclusively to the parietal endoderm following blastocyst injection. The above findings lend no support to a recent proposal that parietal and visceral endoderm are derived from different populations of inner cell mass cells. Rather, they suggest that the two extraembryonic endoderm layers originate from a common pool of primitive endoderm cells whose direction of differentiation depends on their interactions with non-endodermal cells.

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Lineage Establishment and Progression within the Inner Cell Mass of the Mouse Blastocyst Requires FGFR1 and FGFR2

Developmental Cell ◽

10.1016/j.devcel.2017.05.003 ◽

2017 ◽

Vol 41 (5) ◽

pp. 496-510.e5 ◽

Cited By ~ 68

Author(s):

Minjung Kang ◽

Vidur Garg ◽

Anna-Katerina Hadjantonakis

Keyword(s):

Inner Cell Mass ◽

Cell Mass ◽

Mouse Blastocyst

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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

10.21203/rs.3.rs-122470/v1 ◽

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.

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The transcription factor GATA6 is essential for early extraembryonic development

Development ◽

10.1242/dev.126.4.723 ◽

1999 ◽

Vol 126 (4) ◽

pp. 723-732 ◽

Cited By ~ 11

Author(s):

M. Koutsourakis ◽

A. Langeveld ◽

R. Patient ◽

R. Beddington ◽

F. Grosveld

Keyword(s):

Transcription Factor ◽

Inner Cell Mass ◽

Marker Gene ◽

Cell Mass ◽

Blastocyst Stage ◽

Gene Coding ◽

Parietal Endoderm ◽

Reichert's Membrane ◽

Second Wave ◽

Beta Galactosidase

The gene coding for the murine transcription factor GATA6 was inactivated by insertion of a beta-galactosidase marker gene. The analysis of heterozygote GATA6/lacZ mice shows two inductions of GATA6 expression early in development. It is first expressed at the blastocyst stage in part of the inner cell mass and in the trophectoderm. The second wave of expression is in parietal endoderm (Reichert's membrane) and the mesoderm and endoderm that form the heart and gut. Inactivation leads to a lethality shortly after implantation (5.5 days postcoitum). Chimeric experiments show this to be caused by an indirect effect on the epiblast due to a defect in an extraembryonic tissue.

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An investigation of inner cell mass and trophoblast tissues following their isolation from the mouse blastocyst

Development ◽

10.1242/dev.28.2.279 ◽

1972 ◽

Vol 28 (2) ◽

pp. 279-312

Author(s):

R. L. Gardner ◽

M. H. Johnson

Keyword(s):

Inner Cell Mass ◽

Cell Mass ◽

Mouse Blastocyst ◽

Or Groups ◽

Ectoplacental Cone

1. Inner cell mass (ICM) and trophoblast were isolated from 3½-day post-coitum mouse blastocysts by microsurgery. 2. Trophoblastic fragments formed vesicles in culture but did not aggregate with other such fragments. They proved as effective as intact blastocysts in inducing decidua in recipient uteri, but thereafter failed to proliferate. 3. Isolated ICMs remained as solid balls of cells that readily aggregated in pairs or groups in culture but failed to induce implantation changes in receptive uteri. 4. Possible explanations for the failure of isolated trophoblast to proliferate after implantation are discussed. It is argued that presence of ICM tissue is necessary for trophoblast proliferation, and suggested that the ICM exerts its effect by controlling development of the ectoplacental cone.

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