Berberine alleviates LPS-induced apoptosis, oxidation, and skewed lineages during mouse preimplantation development

2022 ◽  
Author(s):  
Xiaosu Miao ◽  
Wei Cui
Keyword(s):  
Inner Cell Mass ◽  
Polycystic Ovary ◽  
Cell Mass ◽  
Cell Lineage ◽  
Culture Conditions ◽  
Preimplantation Development ◽  
Embryonic Cells ◽  
Preimplantation Embryo Development ◽  
New Treatment ◽  
Induced Apoptosis

Abstract Female infertility is a heterogeneous disorder with a variety of complex causes, including inflammation and oxidative stress, which are also closely associated with the pathogenesis of Polycystic Ovary Syndrome (PCOS). As a new treatment for PCOS, berberine (BER), a natural compound from Berberis, has been clinically applied recently. However, the mechanisms underlying the association between BER and embryogenesis are still largely unknown. In this study, effects of BER on preimplantation development was evaluated by using both normal and inflammatory culture conditions induced by lipopolysaccharide (LPS) in the mouse. Our data first suggest that BER itself (25 nM) does not affect embryo quality or future developmental potency, moreover, it can effectively alleviate LPS-induced embryonic damage by mitigating apoptosis via ROS−/caspase-3-dependent pathways and by suppressing pro-inflammatory cytokines via inhibition of NF-κB signaling pathway during preimplantation embryo development. In addition, skewed cell lineage specification in inner cell mass (ICM) and primitive endoderm (PE) caused by LPS can also be successfully rescued with BER. In summary, these findings for the first time demonstrate the non-toxicity of low doses of BER and its anti-apoptotic and anti-oxidative properties on embryonic cells during mammalian preimplantation development.

Effect of culture conditions on diploid to giant-cell transformation in postimplantation mouse trophoblast

Development ◽  
1981 ◽  
Vol 62 (1) ◽  
pp. 217-227
Author(s):  
J. Rossant ◽  
W. Tamura-Lis
Keyword(s):  
Giant Cell ◽  
Cell Transformation ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Cell Formation ◽  
Culture Conditions ◽  
Embryonic Cells ◽  
Giant Cell Formation ◽  
Ectoplacental Cone

Diploid extraembryonic ectoderm and ectoplacental cone from the 7·5-day mouse embryo were grown in vitro under a variety of culture conditions in an attempt to discover conditions which maintain trophoblast in a diploid state and prevent giant-cell formation. It was found that maintenance of tissue integrity was not enough to keep the tissues dividing and diploid, but that the presence of inner-cell-mass derivatives did have some effect. This effect was only apparent when trophoblast cells were entirely enclosed by embryonic tissues. Monolayers of embryonic or embryonal carcinoma cells did not prevent giant-cell formation. Diploid extraembryonic ectoderm and ectoplacental cone responded differently: ectoplacental cells eventually formed trophoblast giant cells even when enclosed by embryonic cells whereas extraembryonic ectoderm cells apparently could be maintained in a diploid condition. This and other differences in properties between extraembryonic ectoderm and ectoplacental cone are discussed with reference to a new model for the postimplantation trophoblast lineage in the mouse.

scholarly journals 50SPATIAL EXPRESSION OF OCT4 IS NORMALLY REGULATED IN PREIMPLANTATION STAGE BOVINE SOMATIC CELL CLONES

2004 ◽  
Vol 16 (2) ◽  
pp. 147
Author(s):  
S. Kurosaka ◽  
S. Eckardt ◽  
M.K. Friez ◽  
N.A. Leu ◽  
R. Reinbold ◽  
...  
Keyword(s):  
Spatial Distribution ◽  
Inner Cell Mass ◽  
Marker Gene ◽  
Cell Mass ◽  
Calf Serum ◽  
Ectopic Expression ◽  
Cell Lineage ◽  
Embryonic Cells ◽  
Oct4 Protein

Oct4 is a member of the POU family of transcription factors and is expressed in blastomeres, pluripotent embryonic cells and the germ cell lineage. In the mouse blastocyst, expression of Oct4 becomes restricted to the inner cell mass (ICM). In bovine, Oct4 has not been considered to be a marker for pluripotency because Oct4 protein is expressed in both the ICM and the trophectoderm (TE) of bovine blastocysts (van Eijk MJT et al., 1999 Biol. Reprod. 60, 1093–1103; Kirchhof N et al., 2000 Biol. Reprod. 63, 1698–1705). Oct4 has been used as a marker gene for nuclear reprogramming in cloned embryos. Aberrant spatial distribution and levels of Oct4 have been observed in the majority of mouse clone blastocysts and blastocyst outgrowths (Boiani M et al., 2002 Genes Dev. 16, 1209–1219), indicating reprogramming failure of mouse clones. Lack of or abnormal Oct4 protein expression was also observed in cloned monkey embryos (Mitalipov SM et al., 2003 Biol. Reprod. 68 (suppl 1), 159). The spatial distribution of Oct4 mRNA and protein in bovine clones has not been reported. Bovine oocytes were obtained from a commercial supplier (BOMED, Inc., Madison, WI, USA), and were matured in vitro. Enucleated oocytes were fused with fibroblasts from ear skin and then treated with 10μgmL−1 cycloheximide and 1.25μgmL−1 cytochalasin D for 6h. Embryos were cultured in SOF supplemented with 1% fetal calf serum (FCS) at 39°C under 5% CO2, 5% O2 and 90% N2 for 2 days. At Day 2, embryos were transferred to SOF supplemented with 5% FCS and cultured under the same conditions until Day 7. Blastocysts were analyzed at Day 7. Oct4 mRNA expression was visualized by whole-mount in situ hybridization using a bovine Oct4-specific antisense riboprobe. Oct4 protein was detected by immunocytochemistry. Control embryos were produced by IVF and were cultured under the same conditions to the blastocyst stage (Day 7). We found that Oct4 mRNA signal was restricted to the ICM in bovine blastocysts. Bovine clones were not different from control embryos in that distribution of Oct4 mRNA signal was typically restricted to the ICM (14 of 16). In contrast to our previous report on mouse clones (Boiani M et al., 2002 Genes Dev. 16, 1209–1219), ectopic expression of Oct4 mRNA in the TE was rarely detected in bovine clones (2 of 16). Distribution of Oct4 protein was also similar between clones and controls with distribution in both the ICM and TE (clones: 9 of 9; controls: 9 of 9). It is unclear why defects in Oct4 distribution should differ between bovine and other species tested including monkey (Mitalipov SM et al., 2003 Biol. Reprod. 68 (suppl 1), 159–160); however, the higher rate of normal Oct4 distribution is consistent with the generally higher rates of postimplantation development of bovine clones (Shi W et al., 2003 Differentiation 71, 91–113).

scholarly journals Cell fate clusters in ICM organoids arise from cell fate heredity and division: a modelling approach

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Tim Liebisch ◽  
Armin Drusko ◽  
Biena Mathew ◽  
Ernst H. K. Stelzer ◽  
Sabine C. Fischer ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Cell Division ◽  
Cell Fate ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Cell Lineage ◽  
Cell Fate Decision ◽  
Cell Fate Decisions ◽  
Chemical Signalling ◽  
Fate Decision

AbstractDuring the mammalian preimplantation phase, cells undergo two subsequent cell fate decisions. During the first decision, the trophectoderm and the inner cell mass are formed. Subsequently, the inner cell mass segregates into the epiblast and the primitive endoderm. Inner cell mass organoids represent an experimental model system, mimicking the second cell fate decision. It has been shown that cells of the same fate tend to cluster stronger than expected for random cell fate decisions. Three major processes are hypothesised to contribute to the cell fate arrangements: (1) chemical signalling; (2) cell sorting; and (3) cell proliferation. In order to quantify the influence of cell proliferation on the observed cell lineage type clustering, we developed an agent-based model accounting for mechanical cell–cell interaction, i.e. adhesion and repulsion, cell division, stochastic cell fate decision and cell fate heredity. The model supports the hypothesis that initial cell fate acquisition is a stochastically driven process, taking place in the early development of inner cell mass organoids. Further, we show that the observed neighbourhood structures can emerge solely due to cell fate heredity during cell division.

scholarly journals Cell lineage allocation in equine blastocysts produced in vitro under varying glucose concentrations

Reproduction ◽  
2015 ◽  
Vol 150 (1) ◽  
pp. 31-41 ◽  
Author(s):  
Young-Ho Choi ◽  
Pablo Ross ◽  
Isabel C Velez ◽  
B Macías-García ◽  
Fernando L Riera ◽  
...  
Keyword(s):  
High Glucose ◽  
Culture Medium ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Cell Lineage ◽  
Primitive Endoderm ◽  
Blastocyst Development ◽  
Embryo Culture Medium ◽  
Effect Of Glucose

Equine embryos developin vitroin the presence of high glucose concentrations, but little is known about their requirements for development. We evaluated the effect of glucose concentrations in medium on blastocyst development after ICSI. In experiment 1, there were no significant differences in rates of blastocyst formation among embryos cultured in our standard medium (DMEM/F-12), which contained >16 mM glucose, and those cultured in a minimal-glucose embryo culture medium (<1 mM; Global medium, GB), with either 0 added glucose for the first 5 days, then 20 mM (0-20) or 20 mM for the entire culture period (20-20). In experiment 2, there were no significant differences in the rates of blastocyst development (31–46%) for embryos cultured in four glucose treatments in GB (0-10, 0-20, 5-10, or 5-20). Blastocysts were evaluated by immunofluorescence for lineage-specific markers. All cells stained positively forPOU5F1. An inner cluster of cells was identified that included presumptive primitive endoderm cells (GATA6-positive) and presumptive epiblast (EPI) cells. The 5-20 treatment resulted in a significantly lower number of presumptive EPI-lineage cells than the 0-20 treatment did.GATA6-positive cells appeared to be allocated to the primitive endoderm independent of the formation of an inner cell mass, as was previously hypothesized for equine embryos. These data demonstrate that equine blastocyst development is not dependent on high glucose concentrations during early culture; rather, environmental glucose may affect cell allocation. They also present the first analysis of cell lineage allocation inin vitro-fertilized equine blastocysts. These findings expand our understanding of the factors that affect embryo development in the horse.

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

Development ◽  
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.

P–516 Evidence for self-correction in preimplantation embryos by comparative molecular karyotyping of blastocoele fluid, trophectoderm and inner cell mass

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
D Zhigalina ◽  
N Skryabin ◽  
O Kanbekova ◽  
V Artyukhova ◽  
A Svetlakov ◽  
...  
Keyword(s):  
Inner Cell Mass ◽  
Cell Mass ◽  
Morphological Characteristics ◽  
Preimplantation Embryos ◽  
Molecular Karyotyping ◽  
Embryonic Cells ◽  
Developmental Potential ◽  
Positive Correlation ◽  
Registration Number ◽  
Molecular Karyotype

Abstract Study question Does the molecular karyotype of the cell-free DNA (cfDNA) from the blastocyst fluid (BF) can predict the efficiency of self-correction of karyotype of preimplantation embryo? Summary answer Detection of aneuploidies in the BF potentially can point out on effective self-correction of blastocyst karyotype and consequently on high developmental potential of mosaic embryos. What is known already Correction of aneuploidies in the preimplantation embryos can be provided by several mechanisms, including apoptosis. The predominant death of aneuploid cells was demonstrated in mouse embryos (Bolton, 2016). A positive correlation was also shown between the concentration of cfDNA from the BF of human blastocyst and the morphology of the embryo, as well as between the activity of caspase–3 and the concentration of cfDNA (Rule, 2018). The incidence of failed amplification after WGA being significantly higher among euploid blastocysts (Magli, 2019). The capacity of abnormal cells extruding into the BF would be related to the embryo development potential (Gianaroli, 2019). Study design, size, duration This is a prospective observational study of thirty-one Day 5 human blastocysts. Cryopreserved blastocysts were received after treatment cycles at the IVF Center with informed consent obtained from couples. The average age of 15 women was 32.25±5 years. The morphological characteristics of blastocysts were estimated in accordance with the Gardner classification (Gardner, Schoolcraft, 1999). The procedure of BF aspiration and trophectoderm (TE) and ICM cells separation of the blastocysts was previously described (Tsuiko, 2018). Participants/materials, setting, methods WGA was performed by PicoPLEX kit (Rubicon Genomics, USA) or REPLI-g Mini kit (Qiagen) according to manufacturer’s protocols. The DNA of the BF, ICM and TE were analyzed separately using cCGH, aCGH and NGS. SurePrint G3 Human CGH Microarrays (8x60K, Agilent Technologies) were used according to the manufacturer’s recommendations. Image analysis was done using ISIS (v.5.5) (Metasystems) and Agilent CytoGenomics Software (v.3). VeriSeq™ PGS Kit - MiSeq® System (Illumina) was used for NGS. Main results and the role of chance Molecular karyotypes of all three samples - BF, ICM and TE, were obtained for 23 (74.2%) blastocysts. A correlation between the woman’s age and the number of aneuploidies in cfDNA (p = 0.0009) was found. A positive correlation may indicate that the number of aneuploidies in the embryonic cells increases with the age of a woman, however, the embryonic karyotype undergoes self-correcting through the elimination of aneuploid cells. It was noted that well-developing blastocysts (groups 4–5, according to Gardner’s classification) had fewer aneuploidies in ICM (p = 0.0141) and TE (p = 0.0436). In contrast, there was a tendency to an increase in the number of aneuploidies in the BF during blastocysts transition from stage 3 to 5 (p = 0.3542). We assessed the relationship between the number of aneuploidies in groups of blastocysts with different characteristics of ICM (groups “A” and “B” according to Gardner’s classification). These groups significantly differ in the number of aneuploidies in cfDNA (p = 0.0352), although the statistically significant differences between the number of aneuploidies in ICM (p = 0.5992) and in TE (p = 0.5934) was not detected. Thus, higher-quality embryos in terms of ICM morphology contain more abnormalities in the BF, since in this group the elimination of aneuploid cells is more efficient. Limitations, reasons for caution The number of embryos is limited in this study. More comprehensive studies are required to confirm the observed tendency. Wider implications of the findings: Aneuploid cells elimination can be a cause of increasing cfDNA concentration in the BF, which may be a marker of the viability of mosaic embryos when it is necessary to decide on mosaic embryo transfer. This study was supported by the RFBR (15–04–08265) and by the RSF (20–74–00064). Trial registration number Not applicable

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.

Metabolic co-operation between embryonic and embryonal carcinoma cells of the mouse

Development ◽  
1979 ◽  
Vol 54 (1) ◽  
pp. 263-275
Author(s):  
Stephen J. Gaunt ◽  
Virginia E. Papaioannou
Keyword(s):  
Embryonal Carcinoma ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Embryonic Cells ◽  
Cone Cells ◽  
Ec Cells ◽  
Hypoxanthine Guanine Phosphoribosyltransferase ◽  
Early Mouse ◽  
Intercellular Exchange ◽  
Embryonic Ectoderm

Mouse embryonal carcinoma (EC) cells form permeable junctions at their homotypic cell-to-cell contacts which permit intercellular exchange of metabolites (metabolic co-operation). Hooper & Slack (1977) showed how this exchange could be detected by autoradiography as the transfer of [3H]nucleotides between PCI3 (a pluripotential EC line) and PCI 3- TG8 (a variant of PC13 which is deficient in hypoxanthine guanine phosphoribosyltransferase). We now show that cells taken from several different tissues of early mouse embryos, that is, from the morula, the inner cell mass of the blastocyst, and the endoderm, mesoderm and embryonic ectoderm of the 8th day egg cylinder, are able to serve as donors of [3H] ucleotides to PC13TG8. In contrast, trophectodermal cells of cultured blastocysts, and the trophectodermal derivatives in the 8th day egg cylinder, that is, extra-embryonic ectoderm and ectoplacental cone cells, showed little or no metabolic co-operation with PC13TG8. With reference to some common properties of EC and embryonic cells, we suggest how our findings may provide insight into cell-to-cell interactions in the early mouse embryo.

Developmental expression and cellular localization of glucose transporter molecules during mouse preimplantation development

Development ◽  
1992 ◽  
Vol 115 (1) ◽  
pp. 305-312
Author(s):  
M. Aghayan ◽  
L.V. Rao ◽  
R.M. Smith ◽  
L. Jarett ◽  
M.J. Charron ◽  
...  
Keyword(s):  
Western Blot ◽  
Cellular Localization ◽  
Glucose Transporter ◽  
Developmental Stages ◽  
Molecular Level ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Blastocyst Stage ◽  
Preimplantation Development ◽  
Developmental Expression

Two general mechanisms mediate glucose transport, one is a sodium-coupled glucose transporter found in the apical border of intestinal and kidney epithelia, while the other is a sodium-independent transport system. Of the latter, several facilitated transporters have been identified, including GLUT1 (erythrocyte/brain), GLUT2 (liver) and GLUT4 (adipose/muscle) isoforms. In this study, we used Western-blot analysis and high resolution immunoelectron microscopy (IEM) to investigate the stage-related expression and cellular localization of GLUT1, 2 and 4. The Western blot results demonstrate that GLUT1 is detectable in the oocyte and throughout preimplantation development. GLUT2 isoforms were not detectable until the blastocyst stage, while the GLUT4 isoform was undetectable in the oocyte through blastocyst stages. The present findings confirm previous studies at the molecular level which demonstrated that mRNAs encoding the same GLUT isoforms are detectable at corresponding developmental stages. GLUT1 and GLUT2 display different cellular distributions at the blastocyst stage as shown by IEM studies. GLUT1 has a widespread distribution in both trophectoderm and inner cell mass cells, while GLUT2 is located on trophectoderm membranes facing the blastocyst cavity. This observation suggests a different functional significance for these isoforms during mouse preimplantation development.

CSF-1 and mouse preimplantation development in vitro

Development ◽  
1995 ◽  
Vol 121 (5) ◽  
pp. 1333-1339 ◽  
Author(s):  
P. Bhatnagar ◽  
V.E. Papaioannou ◽  
J.D. Biggers
Keyword(s):  
Inner Cell Mass ◽  
Cell Mass ◽  
Preimplantation Development ◽  
High Rate ◽  
Colony Stimulating Factor ◽  
Trophoblast Cells ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor

The effects of macrophage colony stimulating factor on the development of the zygote to the blastocyst stage of an outbred strain of mouse have been studied in KSOM, an improved medium that supports a high rate of in vitro development. Macrophage colony stimulating factor accelerates the formation of the blastocyst cavity by day 4 (96 hours post-hCG). It also increases overall embryonic cell number through a differential increase in the number of trophoblast cells, with no significant effect on the number of inner cell mass cells. By day 5 of culture (120 hours post-hCG), colony stimulating factor-treated embryos have about 20 more trophoblast cells than control embryos, an increase of about 30 percent of the total number of cells in a control blastocyst. The maximum response of embryos was obtained at a concentration around 540 U ml-1 colony stimulating factor (identical to 918 Stanley units ml-1), and the cytokine can produce the same effects even if it is present in the medium for only part of the culture period. This in vitro stimulation of preimplantation development with macrophage colony stimulating factor is compatible with continued normal fetal development in vivo.

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