scholarly journals Enzyme analysis of mouse extra-embryonic tissues

Development ◽  
1979 ◽  
Vol 52 (1) ◽  
pp. 127-139
Author(s):  
Michael I. Sherman ◽  
Sui Bl Atienza-Samols
Keyword(s):  
Cell Types ◽  
Embryonic Cell ◽  
Enzyme Analysis ◽  
Visceral Endoderm ◽  
Fetal Circulation ◽  
Embryonic Tissues ◽  
Parietal Endoderm ◽  
Reichert's Membrane ◽  
Catabolic Enzyme ◽  
Alkaline And Acid Phosphatases

We have separated for enzyme analysis the following layers that surround the conceptus at midgestation: decidua, trophoblast, parietal endoderm (including Reichert's membrane), visceral endoderm, yolk-sac mesoderm and amnion. Measurement of several catabolic enzyme activities (N-acetyl-β,D-hexosaminidase, β-glucuronidase, alkaline and acid phosphatases and non-specific esterases) in these tissues indicates that they are biochemically distinct, perhaps reflecting the different functions that they perform in providing the embryo proper with a desirable environment for differentiation and development. Our studies also provide an example of how visceral endoderm cells can effectively block passage of maternal macromolecules (in this case a serum esterase) in the fetal circulation. Finally, since there is often difficulty in distinguishing among early embryonic and extra-embryonic cell types produced in teratocarcinoma cultures, we have considered how our observations might be of use in this respect, particularly in discriminating between visceral and parietal endoderm.

Cell differentiation in isolated inner cell masses of mouse blastocysts in vitro: onset of specific gene expression

Development ◽  
1979 ◽  
Vol 53 (1) ◽  
pp. 367-379
Author(s):  
Marie Dziadek
Keyword(s):  
Gene Expression ◽  
Cell Differentiation ◽  
Biochemical Marker ◽  
Cell Types ◽  
The Other ◽  
Specific Gene ◽  
Visceral Endoderm ◽  
Secondary Layer ◽  
Parietal Endoderm

Inner cell masses (ICMs) were isolated by immunosurgery from giant blastocysts formedby the aggregation of three morulae. A layer of endoderm cells formed on the outer surface of these primary ICMs in vitro. When this layer was removed by immunosurgery, a secondary endoderm layer formed. Alphafetoprotein (AFP) was used as a biochemical marker tocharacterize visceral endoderm formation in these cultured ICMs. The immunoperoxidase reaction on sections of ICMs cultured for intervals up to 120 h in vitro showed that someprimary endoderm cells contained AFP, but these were always in the minority. The secondary endoderm layer, on the other hand, was composed of predominantly AFP-positive cells.Itis concluded that the primary endoderm contains mainly parietal endoderm cells, while the secondary layer contains visceral endoderm cells. A model is proposed for the consecutive differentiation of parietal and visceral endoderm cell types from the ICM of mouseblastocysts.

scholarly journals POSTNATAL EXTRA-EMBRYONIC TISSUES AS A SOURCE OF MULTIPLE CELL TYPES FOR REGENERATIVE MEDICINE APPLICATIONS

2017 ◽  
Vol 39 (3) ◽  
pp. 186-190 ◽  
Author(s):  
O S Gubar ◽  
A I Rodnichenko ◽  
R G Vasylie ◽  
A V Zlatska ◽  
D O Zubov
Keyword(s):  
Regenerative Medicine ◽  
Umbilical Cord ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Umbilical Vein ◽  
Cell Types ◽  
Population Doubling ◽  
Embryonic Tissues ◽  
Chorionic Membrane ◽  
Multiple Cell

Aim: We aimed to isolate and characterize the cell types which could be obtained from postnatal extra-embryonic tissues. Materials and Methods: Fresh tissues (no more than 12 h after delivery) were used for enzymatic or explants methods of cell isolation. Obtained cultures were further maintained at 5% oxygen. At P3 cell phenotype was assessed by fluorescence-activated cell sorting, population doubling time was calculated and the multilineage differentiation assay was performed. Results: We have isolated multiple cell types from postnatal tissues. Namely, placental mesenchymal stromal cells from placenta chorionic disc, chorionic membrane mesenchymal stromal cells (ChM-MSC) from free chorionic membrane, umbilical cord MSC (UC-MSC) from whole umbilical cord, human umbilical vein endothelial cells (HUVEC) from umbilical vein, amniotic epithelial cells (AEC) and amniotic MSC (AMSC) from amniotic membrane. All isolated cell types displayed high proliferation rate together with the typical MSC phenotype: CD73+CD90+CD105+CD146+CD166+CD34-CD45-HLA-DR-. HUVEC constitutively expressed key markers CD31 and CD309. Most MSC and AEC were capable of osteogenic and adipogenic differentiation. Conclusion: We have shown that a wide variety of cell types can be easily isolated from extra-embryonic tissues and expanded ex vivo for regenerative medicine applications. These cells possess typical MSC properties and can be considered an alternative for adult MSC obtained from bone marrow or fat, especially for allogeneic use.

scholarly journals Spatiotemporal sequence of mesoderm and endoderm lineage segregation during mouse gastrulation

2020 ◽  
Author(s):  
Simone Probst ◽  
Sagar ◽  
Jelena Tosic ◽  
Carsten Schwan ◽  
Dominic Grün ◽  
...  
Keyword(s):  
Mouse Embryo ◽  
Primitive Streak ◽  
Cell Types ◽  
Complete Separation ◽  
Embryonic Cell ◽  
Spatiotemporal Pattern ◽  
Cell Lineages ◽  
Dependent Cell ◽  
Summary Statement ◽  
Germ Layer Formation

AbstractAnterior mesoderm (AM) and definitive endoderm (DE) progenitors represent the earliest embryonic cell types that are specified during germ layer formation at the primitive streak (PS) of the mouse embryo. Genetic experiments indicate that both lineages segregate from Eomes expressing progenitors in response to different NODAL signaling levels. However, the precise spatiotemporal pattern of the emergence of these cell types and molecular details of lineage segregation remain unexplored. We combined genetic fate labeling and imaging approaches with scRNA-seq to follow the transcriptional identities and define lineage trajectories of Eomes dependent cell types. All cells moving through the PS during the first day of gastrulation express Eomes. AM and DE specification occurs before cells leave the PS from discrete progenitor populations that are generated in distinct spatiotemporal patterns. Importantly, we don’t find evidence for the existence of progenitors that co-express markers of both cell lineages suggesting an immediate and complete separation of AM and DE lineages.Summary statementCells lineages are specified in the mouse embryo already within the primitive streak where Mesp1+ mesoderm and Foxa2+ endoderm are generated in a spatial and temporal sequence from unbiased progenitors.

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)

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

Development ◽  
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.

Development to term of mouse androgenetic aggregation chimeras

Development ◽  
1991 ◽  
Vol 113 (4) ◽  
pp. 1325-1333 ◽  
Author(s):  
J.R. Mann ◽  
C.L. Stewart
Keyword(s):  
Mouse Strain ◽  
Es Cells ◽  
Embryonic Stem ◽  
Cell Types ◽  
Considerable Improvement ◽  
Es Cell ◽  
Partial Explanation ◽  
Developmental Potential ◽  
Skeletal Abnormalities ◽  
Embryonic Tissues

Diploid androgenetic eggs contain two sperm-derived genomes, and only rarely develop to the early somite stage. Also, previous studies have indicated that androgenetic eggs cannot be rescued in aggregation chimeras beyond embryonic stages. Paradoxically, in blastocyst injection chimeras made with androgenetic embryonic stem (ES) cells of the 129/Sv strain, we previously obtained considerable improvement in developmental potential. Although considerable death occurred in utero, overtly normal chimeric fetuses and occasional postnatal chimeras that developed skeletal abnormalities were observed. Consequently, we have re-evaluated the developmental potential of androgenetic aggregation chimeras utilizing androgenetic eggs of the 129/Sv strain, and of the BALB/c and CD-1 strains for comparison. Regardless of strain, androgenetic aggregation chimeras were generally more inviable than previously observed with androgenetic ES cell chimeras, and often the embryoproper was abnormal even when an androgenetic contribution was detected only in the extra-embryonic membranes. This is at least a partial explanation of the greater viability of androgenetic ES cell chimeras, as ES cells do not colonize significantly certain extra-embryonic tissues. Nevertheless, in the 129/Sv strain, occasional development of chimeras to term was obtained, and one chimera that survived postnatally developed identical skeletal abnormalities to those observed previously in androgenetic ES cell chimeras. This result demonstrates that at least one example of paternal imprinting is faithfully conserved in androgenetic ES cells. Also, the postnatal chimerism shows that androgenetic eggs can give rise to terminally differentiated cell types, and are therefore pluripotent. In contrast, only possibly one BALB/c and no CD-1 androgenetic aggregation chimeras developed to term. Therefore, the developmental potential of androgenetic aggregation chimeras is to some extent dependent on mouse strain.

Cell contacts and sorting out in vivo: the behaviour of some embryonic tissues implanted into the developing chick wing

Development ◽  
1978 ◽  
Vol 48 (1) ◽  
pp. 225-237
Author(s):  
C. Tickle ◽  
M. Goodman ◽  
L. Wolpert
Keyword(s):  
Neural Tube ◽  
Cell Types ◽  
Neural Retina ◽  
Limb Bud ◽  
Embryonic Liver ◽  
Malignant Cells ◽  
Embryonic Tissues ◽  
Mixed Aggregates ◽  
Wing Bud

The interaction of cells from embryonic liver, neural retina and mesonephros with cells from limb-bud mesenchyme has been investigated in vivo by grafting these tissues into the developing chick wing-bud. The implanted cells were in all cases from quail tissue which can be recognized histologically. As embryonic liver and neural tube are tissues that sort externally to limb-bud mesenchyme in mixed aggregates, it would be expected, from a differential adhesiveness hypothesis, that heterotypic adhesions along the borders of graft and host would be favoured over cell-cell adhesions in the graft. No morphological signs of this were evident: rather the grafted cells maximized like-like contacts. The cells of the grafts, including those from control mesenchyme, did not invade into the wing. The results were the same irrespective of whether the graft was a fragment of tissue or a pellet of reaggregated cells. This supports the idea that cells within tissues are not actively moving around and also provides controls for assaying the invasiveness of other cell types, such as malignant cells into the wing.

Mechanisms of Neural Crest Migration

2018 ◽  
Vol 52 (1) ◽  
pp. 43-63 ◽  
Author(s):  
András Szabó ◽  
Roberto Mayor
Keyword(s):  
Neural Crest ◽  
Cell Population ◽  
Epithelial To Mesenchymal Transition ◽  
Neural Crest Cells ◽  
Cell Types ◽  
Embryonic Cell ◽  
Mesenchymal Transition ◽  
Chain Migration ◽  
Mass Migration ◽  
Neural Crest Migration

Neural crest cells are a transient embryonic cell population that migrate collectively to various locations throughout the embryo to contribute a number of cell types to several organs. After induction, the neural crest delaminates and undergoes an epithelial-to-mesenchymal transition before migrating through intricate yet characteristic paths. The neural crest exhibits a variety of migratory behaviors ranging from sheet-like mass migration in the cephalic regions to chain migration in the trunk. During their journey, neural crest cells rely on a range of signals both from their environment and within the migrating population for navigating through the embryo as a collective. Here we review these interactions and mechanisms, including chemotactic cues of neural crest cells’ migration.

scholarly journals Dynamics of trophoblast differentiation in peri-implantation–stage human embryos

2019 ◽  
Vol 116 (45) ◽  
pp. 22635-22644 ◽  
Author(s):  
Rachel C. West ◽  
Hao Ming ◽  
Deirdre M. Logsdon ◽  
Jiangwen Sun ◽  
Sandeep K. Rajput ◽  
...  
Keyword(s):  
Cell Types ◽  
Migration And Invasion ◽  
Human Embryos ◽  
Main Body ◽  
Hormone Production ◽  
Embryonic Tissues ◽  
Maternal Immune System ◽  
Placental Hormone ◽  
First Contact ◽  
Implantation Period

Single-cell RNA sequencing of cells from cultured human blastocysts has enabled us to define the transcriptomic landscape of placental trophoblast (TB) that surrounds the epiblast and associated embryonic tissues during the enigmatic day 8 (D8) to D12 peri-implantation period before the villous placenta forms. We analyzed the transcriptomes of 3 early placental cell types, cytoTB (CTB), syncytioTB (STB), and migratoryTB (MTB), picked manually from cultured embryos dissociated with trypsin and were able to follow sublineages that emerged from proliferating CTB at the periphery of the conceptus. A unique form of CTB with some features of STB was detectable at D8, while mature STB was at its zenith at D10. A form of MTB with a mixed MTB/CTB phenotype arose around D10. By D12, STB generation was in decline, CTB had entered a new phase of proliferation, and mature MTB cells had begun to move from the main body of the conceptus. Notably, the MTB transcriptome at D12 indicated enrichment of transcripts associated with IFN signaling, migration, and invasion and up-regulation of HLA-C, HLA-E, and HLA-G. The STB, which is distinct from the STB of later villous STB, had a phenotype consistent with intense protein export and placental hormone production, as well as migration and invasion. The studies show that TB associated with human embryos is in rapid developmental flux during peri-implantation period when it must invade, signal robustly to the mother to ensure that the pregnancy continues, and make first contact with the maternal immune system.

In vitro production of Reichert's membrane by mouse embryo-derived parietal endoderm cell lines

1990 ◽  
Vol 191 (2) ◽  
pp. 194-203 ◽  
Author(s):  
Kerry J. Fowler ◽  
Katherine Mitrangas ◽  
Marie Dziadek
Keyword(s):  
Cell Lines ◽  
Mouse Embryo ◽  
In Vitro Production ◽  
Endoderm Cell ◽  
Parietal Endoderm ◽  
Reichert's Membrane ◽  
Vitro Production
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