Gene expression profiles during early differentiation of mouse embryonic stem cells

Mansergh, Fiona C.; Daly, Carl S.; Hurley, Anna; Wride, Michael A.; Hunter, Susan; Evans, Martin

doi:10.1186/1471-213x-9-5

Cited by 41 publications

(34 citation statements)

References 58 publications

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“…By influencing ES cell aggregation via the proposed engineering method and furthering our understanding of EB formation, it may be possible to standardize EB-based protocols for efficient and directed ES cell differentiation (Mansergh et al 2009). Here we characterize engineered ES cell aggregation and EB formation, identifying benefits of the novel 3D culture system.…”

Section: Discussionmentioning

confidence: 99%

Controlled embryoid body formation via surface modification and avidin–biotin cross-linking

et al. 2009

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Cell-cell interaction is an integral part of embryoid body (EB) formation controlling 3D aggregation. Manipulation of embryonic stem (ES) cell interactions could provide control over EB formation. Studies have shown a direct relationship between EB formation and ES cell differentiation. We have previously described a cell surface modification and cross-linking method for influencing cell-cell interaction and formation of multicellular constructs. Here we show further characterisation of this engineered aggregation. We demonstrate that engineering accelerates ES cell aggregation, forming larger, denser and more stable EBs than control samples, with no significant decrease in constituent ES cell viability. However, extended culture C5 days reveals significant core necrosis creating a layered EB structure. Accelerated aggregation through engineering circumvents this problem as EB formation time is reduced. We conclude that the proposed engineering method influences initial ES cell-ES cell interactions and EB formation. This methodology could be employed to further our understanding of intrinsic EB properties and their effect on ES cell differentiation.

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Section: Discussionmentioning

confidence: 99%

Controlled embryoid body formation via surface modification and avidin–biotin cross-linking

et al. 2009

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“…Furthermore, recent studies have reported the in vitro differentiation of germ cells from mouse ESCs [10], teratocarcinoma cells [8], human and mouse bone marrow stromal cells (BMSCs) [9,14]. Principally, two different methods have been reported to induce the differentiation of ESCs into germ cells, namely monolayer differentiation [15] and embryoid body (EB) formation [1–3,6,7,10,16,17]. In this line, Geijsen et al [3] and West et al [2] presented the system that requires the differentiation of murine ESCs into EBs and the subsequent isolation of germ cells by non-quantitative gene expression analyses at days 3–9 of EB differentiation.…”

Section: Introductionmentioning

confidence: 99%

In vitro germ cell differentiation from embryonic stem cells of mice: induction control by BMP4 signalling

2016

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The present study aims to confirm and analyse germ cell-related patterns and specific gene expressions at a very early stage of cell commitment. Following the XY cytogenetic confirmation of the CCE mouse embryonic stem cells (mESCs) line, cells were cultured to form embryoid bodies (EBs). Expression pattern assessment of the mouse vasa homologue (Mvh), Stra8, α6 and β1 integrin genes in ESC and 1–3-day-old EB showed that all genes except α6 integrin were expressed in the ESC. The mean calibration of Mvh, Stra8 and α6 integrin expression significantly increased upon EB formation compared with the ESCs. During mouse embryogenesis, the signalling of bone morphogenetic protein (BMP) is essential for germ-line formation. To investigate its role in germ-line induction in vitro, mESCs were cultured as 1-day-old EB aggregates with BMP4 for 4 days in STO co-culture systems, in the presence and absence of 5 ng/ml BMP4. At the end of the culture period, colony assay (number and diameter) was performed and the viability percentage and proliferation rate was determined. There were no significant statistical differences in the abovementioned criteria between these two groups. Moreover, the expression of Mvh, α6 and β1 integrins, Stra8 and Piwil2 genes was evaluated in co-culture groups. The molecular results of co-culture groups showed higher–but insignificant–Piwil2 and significant α6 integrin expressions in BMP4 treated co-culture systems. These results confirmed that the EB system and the presence of BMP4 in a STO co-culture system improve the differentiation of ESCs to germ cell.

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“…Mouse ES cells have been studied in great detail using such global gene expression profiling. [28][29][30][31][32][33][34] These studies have identified genes important for self-renewal and pluripotency of ES cells, as well as important genes for the formation of germ layers (ectoderm, mesoderm, and endoderm). Analysis of genes specific for ES cell maintenance, germ layers, and hematopoiesis indicated that all three germ layers of embryonic development are generated during bioreactor cultures and that each bioreactor shows a unique gene expression profile, especially for hematopoietic differentiation.…”

Section: Introductionmentioning

confidence: 99%

Unique Differentiation Profile of Mouse Embryonic Stem Cells in Rotary and Stirred Tank Bioreactors

Fridley

Fernandez

et al. 2010

Tissue Engineering Part A

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Embryonic stem (ES)-cell-derived lineage-specific stem cells, for example, hematopoietic stem cells, could provide a potentially unlimited source for transplantable cells, especially for cell-based therapies. However, reproducible methods must be developed to maximize and scale-up ES cell differentiation to produce clinically relevant numbers of therapeutic cells. Bioreactor-based dynamic culture conditions are amenable to large-scale cell production, but few studies have evaluated how various bioreactor types and culture parameters influence ES cell differentiation, especially hematopoiesis. Our results indicate that cell seeding density and bioreactor speed significantly affect embryoid body formation and subsequent generation of hematopoietic stem and progenitor cells in both stirred tank (spinner flask) and rotary microgravity (SyntheconÔ) type bioreactors. In general, high percentages of hematopoietic stem and progenitor cells were generated in both bioreactors, especially at high cell densities. In addition, Synthecon bioreactors produced more sca-1 þ progenitors and spinner flasks generated more c-Kit þ progenitors, demonstrating their unique differentiation profiles. cDNA microarray analysis of genes involved in pluripotency, germ layer formation, and hematopoietic differentiation showed that on day 7 of differentiation, embryoid bodies from both bioreactors consisted of all three germ layers of embryonic development. However, unique gene expression profiles were observed in the two bioreactors; for example, expression of specific hematopoietic genes were significantly more upregulated in the Synthecon cultures than in spinner flasks. We conclude that bioreactor type and culture parameters can be used to control ES cell differentiation, enhance unique progenitor cell populations, and provide means for large-scale production of transplantable therapeutic cells.

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Gene expression profiles during early differentiation of mouse embryonic stem cells

Cited by 41 publications

References 58 publications

Controlled embryoid body formation via surface modification and avidin–biotin cross-linking

Controlled embryoid body formation via surface modification and avidin–biotin cross-linking

In vitro germ cell differentiation from embryonic stem cells of mice: induction control by BMP4 signalling

Unique Differentiation Profile of Mouse Embryonic Stem Cells in Rotary and Stirred Tank Bioreactors

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