A functional analysis of imprinting in parthenogenetic embryonic stem cells

Development ◽  
1994 ◽  
Vol 120 (6) ◽  
pp. 1473-1482 ◽  
Author(s):  
N.D. Allen ◽  
S.C. Barton ◽  
K. Hilton ◽  
M.L. Norris ◽  
M.A. Surani
Keyword(s):  
Stem Cells ◽  
Cell Differentiation ◽  
Embryonic Stem Cells ◽  
Growth Regulation ◽  
Embryonic Stem ◽  
Host Cells ◽  
Imprinted Genes ◽  
Developmental Potential

A detailed analysis of the developmental potential of parthenogenetic embryonic stem cells (PGES) was made in vivo and in vitro, and a comparison was made with the development of cells from parthenogenetic embryos (PG). In vivo, in chimeras with normal host cells (N), PGES cells showed a restricted tissue distribution consistent with that of PG cells, suggesting faithful imprinting in PGES cells with respect to genes involved in lineage allocation and differentiation. Restricted developmental potential was also observed in teratomas formed by ectopic transfer under the kidney capsule. In contrast, the classic phenotype of growth retardation normally observed in PG<==>N chimeras was not seen, suggesting aberrant regulation in PGES cells of genes involved in growth regulation. We also analysed the expression of known imprinted genes after ES cell differentiation. Igf2, H19 and Igf2r were all appropriately expressed in the PGES derived cells following induction of differentiation in vitro with all-trans retinoic acid or DMSO, when compared with control (D3) and androgenetic ES cells (AGES). Interestingly, H19 was found to be expressed at high levels following differentiation of the AGES cells. Due to the unexpected normal growth regulation of PGES<==>N chimeras we also examined Igf2 expression in PGES derived cells differentiated in vivo and found that this gene was still repressed. Our studies show that PGES cells provide a valuable in vitro model system to study the effects of imprinting on cell differentiation and they also provide invaluable material for extensive molecular studies on imprinted genes. In addition, the aberrant growth phenotype observed in chimeras has implications for mechanisms that regulate the somatic establishment and maintenance of some imprints. This is of particular interest as aberrant imprinting has recently been invoked in the etiology of some human diseases.

scholarly journals Full-term development of enucleated mouse oocytes fused with embryonic stem cells from different cell lines

Reproduction ◽  
2001 ◽  
pp. 729-733 ◽  
Author(s):  
T Amano ◽  
Y Kato ◽  
Y Tsunoda
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Cell Lines ◽  
Formation Rate ◽  
Embryonic Stem ◽  
Clear Correlation ◽  
Developmental Potential ◽  
Mouse Oocytes

The developmental potential of enucleated mouse oocytes receiving embryonic stem cells from ten lines with either the same or different genetic backgrounds using the cell fusion method was examined in vitro and in vivo. The development of nuclear-transferred oocytes into blastocysts was high (34-88%). However, there was no clear correlation between development into blastocysts after nuclear transfer and the chimaera formation rate of embryonic stem cells. The development into live young was low (1-3%) in all cell lines and 14 of 19 young died shortly after birth. Most of the live young had morphological abnormalities. Of the five remaining mice, two died at days 23 and 30 after birth, but the other three mice are still active at days 359 (mouse 1) and 338 (mice 4 and 5) after birth, with normal fertility. However, the reasons for the abnormalities and postnatal death of embryonic stem cell-derived mice are unknown.

scholarly journals Developmental potential ofGcn5−/− embryonic stem cells in vivo and in vitro

2007 ◽  
Vol 236 (6) ◽  
pp. 1547-1557 ◽  
Author(s):  
Wenchu Lin ◽  
Geraldine Srajer ◽  
Yvonne A. Evrard ◽  
Huy M. Phan ◽  
Yas Furuta ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Developmental Potential

scholarly journals Dickkopf Wnt signaling pathway inhibitor 1 regulates the differentiation of mouse embryonic stem cells in vitro and in vivo

2015 ◽  
Vol 13 (1) ◽  
pp. 720-730 ◽  
Author(s):  
LIPING OU ◽  
LIAOQIONG FANG ◽  
HEJING TANG ◽  
HAI QIAO ◽  
XIAOMEI ZHANG ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Wnt Signaling ◽  
Signaling Pathway ◽  
Wnt Signaling Pathway ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cells

Derivation of human embryonic stem cell lines after blastocyst microsurgery

2010 ◽  
Vol 88 (3) ◽  
pp. 479-490 ◽  
Author(s):  
Guoliang Meng ◽  
Shiying Liu ◽  
Xiangyun Li ◽  
Roman Krawetz ◽  
Derrick E. Rancourt
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Cell Lines ◽  
Clinical Medicine ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Embryonic Stem ◽  
Hesc Lines

Embryonic stem cells (ESCs) are derived from the inner cell mass (ICM) of the blastocyst. Because of their ability to differentiate into a variety of cell types, human embryonic stem cells (hESCs) provide an unlimited source of cells for clinical medicine and have begun to be used in clinical trials. Presently, although several hundred hESC lines are available in the word, only few have been widely used in basic and applied research. More and more hESC lines with differing genetic backgrounds are required for establishing a bank of hESCs. Here, we report the first Canadian hESC lines to be generated from cryopreserved embryos and we discuss how we navigated through the Canadian regulatory process. The cryopreserved human zygotes used in this study were cultured to the blastocyst stage, and used to isolate ICM via microsurgery. Unlike previous microsurgery methods, which use specialized glass or steel needles, our method conveniently uses syringe needles for the isolation of ICM and subsequent hESC lines. ICM were cultured on MEF feeders in medium containing FBS or serum replacer (SR). Resulting outgrowths were isolated, cut into several cell clumps, and transferred onto fresh feeders. After more than 30 passages, the two hESC lines established using this method exhibited normal morphology, karyotype, and growth rate. Moreover, they stained positively for a variety of pluripotency markers and could be differentiated both in vitro and in vivo. Both cell lines could be maintained under a variety of culture conditions, including xeno-free conditions we have previously described. We suggest that this microsurgical approach may be conducive to deriving xeno-free hESC lines when outgrown on xeno-free human foreskin fibroblast feeders.

230 GENERATION OF PUTATIVE RABBIT EMBRYONIC STEM CELLS

2007 ◽  
Vol 19 (1) ◽  
pp. 231
Author(s):  
S. Wang ◽  
X. Tang ◽  
Y. Niu ◽  
H. Chen ◽  
T. Li ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
High Speed ◽  
Es Cells ◽  
Research Work ◽  
Embryonic Stem ◽  
Morphological Characteristics ◽  
Mouse Escs

The rabbit, as a laboratory animal model, has several advantages in the study of human physiological disorders. In this study, stable putative pluripotent rabbit embryonic stem cells (rESCs) were derived from in vivo-fertilized and in vitro-cultured blastocysts. The rabbit ICMs were obtained by 0.05% trypsin–0.008% EDTA treatment and mechanical separation; the ES-like cell colonies seen several days later. ICM-derived outgrowths which were treated with 5 mg/mL-1 dispase, followed by 0.05% trypsin–0.008% EDTA, were mechanically disaggregated into small clumps and reseeded on MEFs. The putative ES cell lines maintained expression of pluripotent cells markers and normal XY karyotype for long periods of culture (>1 month). The putative rESCs expressed alkaline phosphatase, transcription factor Oct-4, stage-specific embryonic antigens (SSEA-1, SSEA-3, and SSEA-4), and tumor-related antigens (TRA-1-60 and TRA-1-81). The morphological characteristics of the putative ESCs are closer to those of human ESCs; their high speed of proliferation, however, is closer to that of mouse ESCs. Putative rabbit ESCs were induced to differentiate into many cell types including trophoblast cells, similar to primate ESCs, in vitro, and formed teratomas with derivatives of the 3 major germ layers in vivo when injected into SCID mice. Using RT-PCR measurement, but with some differences in ligands and inhibitors, and comparing with human and mouse ESCs, the putative rabbit ESCs expressed similar genes related to pluripotency (Oct-4, Nanog, SOX2, and UTF-1) and similar genes of FGF, WNT, and TGF signaling pathways related to the proliferation and self-renewal. Our further research work showed that TGF beta and FGF pathways cooperate to maintain pluripotency of rabbit ESCs similar to those of human ES cells.

In Vitro Differentiation and In Vivo Mineralization of Osteogenic Cells Derived from Human Embryonic Stem Cells

2004 ◽  
Vol 10 (9-10) ◽  
pp. 1518-1525 ◽  
Author(s):  
Robert C. Bielby ◽  
Aldo R. Boccaccini ◽  
Julia M. Polak ◽  
Lee D.K. Buttery
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
In Vitro Differentiation ◽  
Osteogenic Cells
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