scholarly journals The Combination of Inhibitors of FGF/MEK/Erk and GSK3β Signaling Increases the Number of OCT3/4- and NANOG-Positive Cells in the Human Inner Cell Mass, But Does Not Improve Stem Cell Derivation

2013 ◽  
Vol 22 (2) ◽  
pp. 296-306 ◽  
Author(s):  
Margot Van der Jeught ◽  
Thomas O'Leary ◽  
Sabitri Ghimire ◽  
Sylvie Lierman ◽  
Galbha Duggal ◽  
...  
Keyword(s):  
Stem Cell ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Stem Cell Derivation

Leptin treatment of in vitro cultured embryos increases outgrowth rate of inner cell mass during embryonic stem cell derivation

2019 ◽  
Vol 55 (7) ◽  
pp. 473-481 ◽  
Author(s):  
Ali Cihan Taskin ◽  
Ahmet Kocabay ◽  
Ayyub Ebrahimi ◽  
Sercin Karahuseyinoglu ◽  
Gizem Nur Sahin ◽  
...  
Keyword(s):  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Embryonic Stem ◽  
Stem Cell Derivation

Tracking the progression of the human inner cell mass during embryonic stem cell derivation

2012 ◽  
Vol 30 (3) ◽  
pp. 278-282 ◽  
Author(s):  
Thomas O'Leary ◽  
Björn Heindryckx ◽  
Sylvie Lierman ◽  
David van Bruggen ◽  
Jelle J Goeman ◽  
...  
Keyword(s):  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Embryonic Stem ◽  
Stem Cell Derivation

396 Oct-4 EXPRESSION IN CAT INNER CELL MASS-DERIVED CELLS CULTURED IN MEDIUM WITH DIFFERENT PROTEIN SOURCES

2010 ◽  
Vol 22 (1) ◽  
pp. 354
Author(s):  
T. S. Rascado ◽  
J. F. Lima-Neto ◽  
S. E. R. S. Lorena ◽  
B. W. Minto ◽  
F. C. Landim-Alvarenga
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Inner Cell Mass ◽  
Culture Media ◽  
Cell Mass ◽  
Embryonic Stem ◽  
Domestic Cat ◽  
Santa Cruz

The domestic cat can be used as a biological model for humans because of similarities in some disease and genetically transmitted conditions. Embryonic stem cells might complete nuclear reprogramming more efficiently than somatic cells and, therefore, are potentially useful for increasing interspecific cloning success. The objective of this study was to establish an effective culture system for inner cell mass (ICM)-derived cells in the domestic cat, testing the ability of the ICM to attach to the culture dish and to form embryonic stem cell colonies in the presence of fetal calf serum (FCS) and Knockout serum (KS). Moreover, knowing that the transcription factor Oct-4 is important for the maintenance of pluripotency in human and murine embryonic stem cells, the expression of this factor was evaluated in in vitro-produced blastocyst and in the attached ICM. Domestic cat oocytes were matured, fertilized, and cultured in vitro until the blastocyst stage. The ICM was mechanically isolated (n = 60) using a scalpel blade and transferred to a monolayer of chemically inactivated cat fibroblasts with 10 μg mL-1 mitomicin C. The base culture media (BM) was DMEM/F12 supplemented with nonessential amino acids, glutamine, leukemia inhibitory factor, fibroblast growth factor-2, 2-mercaptoethanol, and antibiotics. Three groups were tested: G1 = BM with 20% FCS (20); G2 = BM with 20% KS (20); G3 = BM with 15% FSC and 5% KS (20). Culture was performed in a 5% CO2 in air incubator at 38.5°C. No statistical difference was observed among groups in relation to ICM attachment (chi-square, P > 0.05). Ninety percent of the ICM presented good adhesion after 3 days of culture and started to grow in all media tested. However, until now, no good colonies were formed. Fifteen blastocysts and 10 attached ICM were fixed in 3% paraformaldehyde and permeabilized in 0.2% triton X-100 in PBS. Subsequently, to block nonspecific binding of the primary antibody, the preadsorption for 2 h at room temperature with OCT4 blocking peptide (sc-8628P, Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used. Samples were incubated with Oct4 antibody (N-19 : sc 8628, Santa Cruz Biotechnology) and with the appropriate secondary antibody (A21431, Invitrogen) and examined by fluorescence microscopy. Oct4 protein was detected both in the ICM and trophoderm cells, and it was distributed in cytoplasm and nuclei. These embryos were also stained with Hoechst 33342. Although further standardization of the culture media is needed, it seems that the KS can be replaced by FCS in cat embryonic stem cell culture. Furthermore, the immunostain of the trophoderm with Oct-4 indicates a difference in the expression of this factor when compared with its expression on human and murine blastocysts. This could be related to in vitro production, or Oct 4 is not a good pluripotency marker for cat embryos and cat embryonic stem cell, consequently. This fact has been noted in goat, bovine, and porcine embryos. Acknowledgment is given to FAPESP.

scholarly journals Stem Cell and Markers

2019 ◽  
Vol 1 (1) ◽  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem Cells ◽  
Adult Stem Cells ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Embryonic Stem ◽  
Stem Cell Markers ◽  
Cell Markers ◽  
Wide Range

Stem cells have the ability to go through various cell divisions and also maintain undifferentiated state. Stem cells are Embryonic (Pluripotent) and adult stem cells. Pluripotent stem cells give rise to all tissues such as ectoderm, mesoderm and endoderm. Embryonic stem cells isolated from inner cell mass of embryo blastocyst. Adult stem cells are also undifferentiated cells present in adult organisms and repair the tissue when damaged occurs but number in less. Adult stem cells are present in bone marrow, adipose tissue, blood and juvenile state umbilical cord and tissue of specific origin like liver, heart, intestine and neural tissue. Embryonic stem cells from blastocyst have the ethical problems and tumorogenecity. These can be identified by flow cytometry. There are wide range of stem cell markers which are useful in identifying them. Most of the pluripotent cell markers are common with tumor cell markers which throws a challenge for certainty.

scholarly journals The Ethical Case against Stem Cell Research

2003 ◽  
Vol 12 (4) ◽  
pp. 372-383 ◽  
Author(s):  
SØREN HOLM
Keyword(s):  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Stem Cell Research ◽  
Human Embryonic Stem Cell ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Embryonic Stem ◽  
Ethical Analysis ◽  
Considerable Debate ◽  
Stem Cell Lines

The possibility of creating human embryonic stem cell lines from the inner cell mass of blastocysts has led to considerable debate about how these scientific developments should be regulated. Part of this debate has focused on the ethical analysis and part on how this analysis should influence policymaking.

scholarly journals Mechanical isolation of the inner cell mass is effective in derivation of new human embryonic stem cell lines

2007 ◽  
Vol 22 (12) ◽  
pp. 3051-3058 ◽  
Author(s):  
S. Strom ◽  
J. Inzunza ◽  
K.-H. Grinnemo ◽  
K. Holmberg ◽  
E. Matilainen ◽  
...  
Keyword(s):  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Cell Lines ◽  
Human Embryonic Stem Cell ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Embryonic Stem ◽  
Stem Cell Lines ◽  
Mechanical Isolation

scholarly journals The post-inner cell mass intermediate: implications for stem cell biology and assisted reproductive technology

2015 ◽  
Vol 21 (5) ◽  
pp. 616-626 ◽  
Author(s):  
Margot Van der Jeught ◽  
Thomas O'Leary ◽  
Galbha Duggal ◽  
Petra De Sutter ◽  
Susana Chuva de Sousa Lopes ◽  
...  
Keyword(s):  
Stem Cell ◽  
Assisted Reproductive Technology ◽  
Cell Biology ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Reproductive Technology ◽  
Stem Cell Biology

scholarly journals Spatially resolved cell polarity proteomics of a human epiblast model

2021 ◽  
Vol 7 (17) ◽  
pp. eabd8407
Author(s):  
Sicong Wang ◽  
Chien-Wei Lin ◽  
Amber E. Carleton ◽  
Chari L. Cortez ◽  
Craig Johnson ◽  
...  
Keyword(s):  
Stem Cell ◽  
Inner Cell Mass ◽  
Basolateral Membrane ◽  
Cell Mass ◽  
Three Dimensional ◽  
Membrane Domains ◽  
Spatially Resolved ◽  
Labeling Approach ◽  
Clathrin Adaptor ◽  
Specific Membrane

Critical early steps in human embryonic development include polarization of the inner cell mass, followed by formation of an expanded lumen that will become the epiblast cavity. Recently described three-dimensional (3D) human pluripotent stem cell–derived cyst (hPSC-cyst) structures can replicate these processes. To gain mechanistic insights into the poorly understood machinery involved in epiblast cavity formation, we interrogated the proteomes of apical and basolateral membrane territories in 3D human hPSC-cysts. APEX2-based proximity bioinylation, followed by quantitative mass spectrometry, revealed a variety of proteins without previous annotation to specific membrane subdomains. Functional experiments validated the requirement for several apically enriched proteins in cyst morphogenesis. In particular, we found a key role for the AP-1 clathrin adaptor complex in expanding the apical membrane domains during lumen establishment. These findings highlight the robust power of this proximity labeling approach for discovering novel regulators of epithelial morphogenesis in 3D stem cell–based models.

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