Growth and differentiation factors of pluripotential stem cells

Heath, John K.; Smith, Austin; Hsu, Li-Wei; Rathjen, Peter D.

doi:10.1242/jcs.1990.supplement_13.8

Cited by 17 publications

(3 citation statements)

References 58 publications

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“…Prompted by observations in murine ES‐cell cultures (M artin 1981; M artin and L ock 1983; H andyside et al 1989), we attempted to culture porcine ES‐cells in conditioned medium, but since none of the conditioned media tested fulfilled our expectations (N eubert 1992), LIF‐ or ESG‐supplemented basic media were used. The homologous growth factors LIF (myeloid Leukaemia Inhibitory Factor) and DIA (Differentiation Inhibiting Activity) are commonly included in murine ESC cultures (W illiams et al 1988; H eath et al 1990; N ichols et al 1990). Their differentiation‐inhibiting effect on pluripotent cells has frequently been reported.…”

Section: Discussionmentioning

confidence: 99%

Identification, isolation and culture of pluripotent cells from the porcine inner cell mass

Ropeter‐Scharfenstein¹,

Neubert²,

Prelle³

et al. 1996

J Animal Breeding Genetics

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Summary This investigation addresses the issue of establishing a porcine embryonic stem cell line. By staining for the intracellular filaments vimentin and keratin in whole‐mount‐preparations it was concluded that the ICM has to be isolated before D9. Although the isolation by way of Ca‐ionophore or immunosurgery was possible, a mechanical approach was preferred. A range of feeder cell layers—both primary and established cell lines—and of conditioned media was found to be less suitable for the culture of ICM cells than was DMEM substituted with LIF or ESG. Eventually a two‐stage culture system was established comprising a 4 day culture of embryos on a three‐dimensional collagen matrix followed by culture of the ICM in a fortified medium supplemented with LIF. With this system, undifferentiated ICM cells could be cultured on average 5.6 and up to 8 passages of 7–10 days each. Attempts to use these cells to make chimaeras or embryo clones are forthcoming. Zusammenfassung Identifizierung, Isolierung und Kultur pluripotenter Zellen der Inneren Zellmasse (ICM) beim Schwein Das Ziel der Arbeit war die Erstellung porciner embryonaler Stammzellinien. Als erstes wurde durch Intermediärfilamentfärbung von ‘whole mount'‐Präparaten mit Anti‐Keratin und Anti‐Vimentin ermittelt, daß eine Isolierung der ICM vor D9 zu erfolgen hat. Obschon dies auf immunologischem Wege oder mit Hilfe von Ca‐Ionophor möglich war, wurde der mechanischen Isolierung der Vorzug gegeben. Die Kultur von ICM‐Zellen in DMEM mit LIF oder ESG war erfolgreicher als auf verschiedenen Feeder layern—sowohl primären als auch etablierten Zellinien — oder in konditionierten Medien. Am geeignetsten erwies sich eine Zwei‐Phasen‐Kultur bestehend aus einer 4‐tägigen Vorkultivierung der Embryonen auf einer dreidimensionalen Kollagen‐Matrix mit anschließender Kultivierung der isolierten ICM in LIF‐haltigem angereicherten DMEM Medium. Damit wurden durchschnittlich 5,6 und maximal 8 Passagen von jeweils 7–10 Tagen erreicht, bis die Vitalität der Zellen erschöpft war. Derzeitige Bemühungen richten sich auf die Verwendung dieser Zellen zur Erstellung von Injektionschimären oder zur Embryoklonierung.

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Section: Discussionmentioning

confidence: 99%

Identification, isolation and culture of pluripotent cells from the porcine inner cell mass

Ropeter‐Scharfenstein¹,

Neubert²,

Prelle³

et al. 1996

J Animal Breeding Genetics

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“…It was also noted that the initial ES cell plating density strongly affects subsequent ES cell/EB differentiation. At cell densities equal or greater than 10,000 cells per ml in CDM, ES cell differentiation was inhibited (Table 3); this effect is presumably due to a factor(s) produced by the ES cells themselves, such as LIF (19). All subsequent differentiation studies used a suspension of 5,000 cells per ml of CDM plus 1 U of LIF per ml per 35-mm-diameter non-tissue culture plate.…”

Section: Es Cells Survive and Differentiate In Cdm In The Absence Of mentioning

confidence: 99%

Evidence for Involvement of Activin A and Bone Morphogenetic Protein 4 in Mammalian Mesoderm and Hematopoietic Development

Johansson¹,

Wiles²

1995

Molecular and Cellular Biology

418

274

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Xenopus in vitro studies have implicated both transforming growth factor ␤ (TGF-␤) and fibroblast growth factor (FGF) families in mesoderm induction. Although members of both families are present during mouse mesoderm formation, there is little evidence for their functional role in mesoderm induction. We show that mouse embryonic stem cells, which resemble primitive ectoderm, can differentiate to mesoderm in vitro in a chemically defined medium (CDM) in the absence of fetal bovine serum. In CDM, this differentiation is responsive to TGF-␤ family members in a concentration-dependent manner, with activin A mediating the formation of dorsoanterior-like mesoderm and bone morphogenetic protein 4 mediating the formation of ventral mesoderm, including hematopoietic precursors. These effects are not observed in CDM alone or when TGF-␤1, -␤2, or -␤3, acid FGF, or basic FGF is added individually to CDM. In vivo, at day 6.5 of mouse development, activin ␤A RNA is detectable in the decidua and bone morphogenetic protein 4 RNA is detectable in the egg cylinder. Together, our data strongly implicate the TGF-␤ family in mammalian mesoderm development and hematopoietic cell formation.

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“…In line with its expression in the inner cell mass and primitive neuroectoderm of early mouse embryos, the Oct4 gene is expressed at high levels in both embryonic stem (ES) cells and embryonal carcinoma (EC) cells (59,81). These cell types are frequently used as in vitro model systems to study early developmental processes (21,25,86). In both EC and ES cells, expression of Oct4 is rapidly down-regulated during retinoic acid (RA)-induced differentiation (45,59,81,83).…”

mentioning

confidence: 99%

Characterization of a negative retinoic acid response element in the murine Oct4 promoter.

Schoorlemmer¹,

Puijenbroek²,

Eijnden³

et al. 1994

Mol. Cell. Biol.

100

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Expression of Oct4 in embryonic stem cells is controlled by a distal upstream stem cell-specific enhancer that is deactivated during retinoic acid (RA)-induced differentiation by an indirect mechanism not involving binding of RA receptors (H. Okazawa, K. Okamoto, F. Ishino, T. Ishino-Kaneko, S. Takeda, Y. Toyoda, M. Muramatsu, and H. Hamada, EMBO J. 10:2997EMBO J. 10: -3005, 1991). Here we report that in RA-treated P19 embryonal carcinoma cells the Oct4 promoter is also subject to negative regulation by RA. The minimal Oct4 promoter sequence mediating repression consists of a promoter-proximal sequence containing a GC-rich SP1 consensus-like sequence and several hormone response element half-sites that can be arranged into direct repeats with different spacing. The GC box binds a nuclear factor that is invariably present in undifferentiated and RA-treated differentiated P19 cells. By contrast, the hormone response element-containing sequence binds factors that are induced following RA treatment. Mutational analysis and competition experiments show that the functional entity binding the RA-induced factor is a direct repeat sequence with a spacing of one nucleotide, previously shown to be a binding site for COUP transcription factors (COUP-TFs). Cotransfected orphan receptors COUP-TF1, ARP-1, and EAR-2 were able to repress the activity of Oct4 promoter-driven reporters in P19 EC cells, albeit with different efficiencies. Furthermore, the negative transcriptional effect of COUP-TFs is dominant over the activating effect of the Oct4 embryonic stem cell-specific enhancer. These results show that negative regulation of Oct4 expression during RA-induced differentiation of embryonic stem cells is controlled by two different mechanisms, including deactivation of the embryonic stem cell-specific enhancer and promoter silencing by orphan nuclear hormone receptors.Among the gene families that play an important role during both vertebrate and invertebrate development is the POU family of transcriptional regulators. Their conserved DNAbinding motif, first recognized among the Pit-1/GHF-1, Octl and Oct2, and Unc83 proteins (27), is involved in binding to the octamer motif (18,95,96), present in the enhancer/ promoter of a variety of eukaryotic genes (77, 85). The Oct family presently consists of a score of members, including Octl (87), Oct2 (56,78), Oct4 (59,68,82), and Oct6 (53,55,88), that, with the exception of Octl, have been implicated in tissue-and stage-specific transcriptional regulation in line with their restricted expression patterns during early embryogenesis and in adult tissues (67,72,79). Expression of Oct4 during early mouse development is restricted to pluripotent stem cells as found in the inner cell mass of blastocyst stage embryos, the pluripotent primitive ectoderm, primordial germ cells, oocytes, and spermatids (68,80,81). In line with its expression in the inner cell mass and primitive neuroectoderm of early mouse embryos, the Oct4 gene is expressed at high levels in both embryonic stem (ES) cells and em...

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Growth and differentiation factors of pluripotential stem cells

Cited by 17 publications

References 58 publications

Identification, isolation and culture of pluripotent cells from the porcine inner cell mass

Identification, isolation and culture of pluripotent cells from the porcine inner cell mass

Evidence for Involvement of Activin A and Bone Morphogenetic Protein 4 in Mammalian Mesoderm and Hematopoietic Development

Characterization of a negative retinoic acid response element in the murine Oct4 promoter.

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