scholarly journals Trophoblast stem cell marker gene expression in inner cell mass-derived cells from parthenogenetic equine embryos

Reproduction ◽  
2011 ◽  
Vol 141 (3) ◽  
pp. 321-332 ◽  
Author(s):  
Joëlle A Desmarais ◽  
Simon-Pierre Demers ◽  
Joao Suzuki ◽  
Simon Laflamme ◽  
Patrick Vincent ◽  
...  
Keyword(s):  
Stem Cell ◽  
Cell Therapy ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Stem Cell Marker ◽  
Large Animal ◽  
Stem Cell Markers ◽  
Trophoblast Stem ◽  
Trophoblast Stem Cell ◽  
Therapy Studies

Although putative horse embryonic stem (ES)-like cell lines have been obtained recently from in vivo-derived embryos, it is currently not known whether it is possible to obtain ES cell (ESC) lines from somatic cell nuclear transfer (SCNT) and parthenogenetic (PA) embryos. Our aim is to establish culture conditions for the derivation of autologous ESC lines for cell therapy studies in an equine model. Our results indicate that both the use of early-stage blastocysts with a clearly visible inner cell mass (ICM) and the use of pronase to dissect the ICM allow the derivation of a higher proportion of primary ICM outgrowths from PA and SCNT embryos. Primary ICM outgrowths express the molecular markers of pluripotency POU class 5 homeobox 1 (POU5F1) and (sex determining region-Y)-box2 (SOX2), and in some cases, NANOG. Cells obtained after the passages of PA primary ICM outgrowths display alkaline phosphatase (AP) activity and POU5F1, SOX2, caudal-related homeobox-2 (CDX2) and eomesodermin (EOMES) expression, but may lose NANOG. Cystic embryoid body-like structures expressing POU5F1, CDX2 and EOMES were produced from these cells. Immunohistochemical analysis of equine embryos reveals the presence of POU5F1 in trophectoderm, primitive endoderm and ICM. These results suggest that cells obtained after passages of primary ICM outgrowths are positive for trophoblast stem cell markers while expressing POU5F1 and displaying AP activity. Therefore, these cells most likely represent trophoblast cells rather than true ESCs. This study represents an important first step towards the production of autologous equine ESCs for pre-clinical cell therapy studies on large animal models.

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.

302 IDENTIFICATION OF 3i TARGET MOLECULES AND THEIR INVOLVEMENT IN PORCINE PLURIPOTENCY NETWORKS

2013 ◽  
Vol 25 (1) ◽  
pp. 298
Author(s):  
G. Pennarossa ◽  
S. Maffei ◽  
M. M. Rahman ◽  
F. Gandolfi ◽  
T. A. L. Brevini
Keyword(s):  
Stem Cell ◽  
Cell Lines ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Embryonic Stem ◽  
Large Animal ◽  
Rt Pcr ◽  
Pluripotent Cell ◽  
Pluripotency Markers ◽  
Target Molecules

Recent studies have shown that the use of specific inhibitors for signalling pathways known to drive murine embryonic stem cell (ESC) differentiation may represent a tool to derive and maintain pluripotent cell lines. The application of this novel approach could provide a new strategy to overcome the limitations still existing for the derivation of ESC in large animal species. These molecules, also known as 3i factors, include CHIR99021 (GSK3 inhibitor), PD173074 (FGF inhibitor) and PD0325901 (MAPK/ERK kinase or MEK inhibitor). However only scattered information are available on the involvement of these pathways in the maintenance of pluripotency in domestic animals. The aim of this study was to investigate the presence of receptors for these inhibitors in porcine inner cell mass (ICM) and to isolate and culture porcine pluripotent lines in a serum-free medium supplemented with the 3i factors, without any additional growth factor. Ovaries were collected at the local abattoir and cumulus–oocyte complexes (COC) were aspirated from antral follicles. In vitro maturation was then performed for 46 h. Frozen–thawed spermatozoa were purified and live spermatozoa were co-cultured with denuded oocytes for 24 h. Putative embryos were cultured in NCSU-23 medium until the blastocyst stage and then subjected to immuno-surgery. Isolated ICM were analysed by RT-PCR. Poly(A)+RNA was extracted using Dynabeads® mRNA DIRECT Micro-kit (Invitrogen, Carlsbad, CA, USA) and immediately reverse-transcribed with Superscript™ II Reverse Transcriptase (Invitrogen). Specific primers were designed for FGF4, FGFR-1, FGFR-2, FGFR-4, GSK3, and MEK genes. The PCR amplified products were sequenced and aligned using ClustalW. The RT-PCR results showed that porcine ICM actively transcribe for GSK3, MEK, FGFR-2, FGFR-4, and FGF4 genes, whereas no signal was detectable for FGFR-1. Based on these observations, IVF-derived ICM were plated onto inactivated STO feeder cells and cultured in N2B27 medium supplemented with 3 µM CHIR99021, 100 nM PD173074, and 0.4 µM PD0325901. Outgrowth formation was monitored and cells were passaged to a new STO monolayer every 7 days, as previously described (Brevini et al. 2010 Stem Cell Rev.). Assessment of pluripotency markers was carried out both by RT-PCR and immunocytochemical analysis at every passage for up to 15 passages. The results obtained indicate that porcine cells cultured in 3i medium, without additional growth factors, expressed pluripotency markers; namely OCT4, NANOG, SOX2, and REX1, preserving their pluripotent state over time. Our data indicate that porcine ICM express 3i factor target molecules. In agreement with this, the use of 3i medium allows the establishment and proliferation of pluripotent cell lines. Together, these findings suggest the involvement of the GSK3, FGF, and MEK pathways in porcine pluripotency network and advocate the use of the 3i medium as an efficient tool for ESC derivation in porcine. Supported by NetLiPS Project ID: 30190629.

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

Lgr5 expression as stem cell marker in human gastric gland and its relatedness with other putative cancer stem cell markers

Gene ◽  
2013 ◽  
Vol 525 (1) ◽  
pp. 18-25 ◽  
Author(s):  
Chen Wu ◽  
Yuanyuan Xie ◽  
Feng Gao ◽  
Yanan Wang ◽  
Yawei Guo ◽  
...  
Keyword(s):  
Stem Cell ◽  
Cancer Stem Cell ◽  
Gastric Gland ◽  
Stem Cell Marker ◽  
Stem Cell Markers ◽  
Cell Marker ◽  
Cell Markers ◽  
Cancer Stem Cell Markers ◽  
Lgr5 Expression

scholarly journals ECM-Dependent HIF Induction Directs Trophoblast Stem Cell Fate via LIMK1-Mediated Cytoskeletal Rearrangement

PLoS ONE ◽  
2013 ◽  
Vol 8 (2) ◽  
pp. e56949 ◽  
Author(s):  
Hwa J. Choi ◽  
Timothy A. Sanders ◽  
Kathryn V. Tormos ◽  
Kurosh Ameri ◽  
Justin D. Tsai ◽  
...  
Keyword(s):  
Stem Cell ◽  
Cell Fate ◽  
Stem Cell Fate ◽  
Cytoskeletal Rearrangement ◽  
Trophoblast Stem ◽  
Trophoblast Stem Cell

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

scholarly journals Connexin31.1 (Gjb5) Deficiency Blocks Trophoblast Stem Cell Differentiation and Delays Placental Development

2014 ◽  
Vol 23 (21) ◽  
pp. 2649-2660 ◽  
Author(s):  
Mark Kibschull ◽  
Keith Colaco ◽  
Elzbieta Matysiak-Zablocki ◽  
Elke Winterhager ◽  
Stephen J. Lye
Keyword(s):  
Stem Cell ◽  
Cell Differentiation ◽  
Stem Cell Differentiation ◽  
Placental Development ◽  
Trophoblast Stem ◽  
Trophoblast Stem Cell
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