scholarly journals Changes in Cell Morphology and Actin Organization in Embryonic Stem Cells Cultured under Different Conditions

Cells ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 2859
Author(s):  
Younes F. Barooji ◽  
Kasper G. Hvid ◽  
Irene Istúriz Petitjean ◽  
Joshua M. Brickman ◽  
Lene B. Oddershede ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Cell Morphology ◽  
Ex Vivo ◽  
Embryonic Stem ◽  
Super Resolution ◽  
Cell Periphery ◽  
Regulatory Structure ◽  
Actin Structure ◽  
Cells Cultured

The cellular cytoskeleton provides the cell with a mechanical rigidity that allows mechanical interaction between cells and the extracellular environment. The actin structure plays a key role in mechanical events such as motility or the establishment of cell polarity. From the earliest stages of development, as represented by the ex vivo expansion of naïve embryonic stem cells (ESCs), the critical mechanical role of the actin structure is becoming recognized as a vital cue for correct segregation and lineage control of cells and as a regulatory structure that controls several transcription factors. Naïve ESCs have a characteristic morphology, and the ultrastructure that underlies this condition remains to be further investigated. Here, we investigate the 3D actin cytoskeleton of naïve mouse ESCs using super-resolution optical reconstruction microscopy (STORM). We investigate the morphological, cytoskeletal, and mechanical changes in cells cultured in 2i or Serum/LIF media reflecting, respectively, a homogeneous preimplantation cell state and a state that is closer to embarking on differentiation. STORM imaging showed that the peripheral actin structure undergoes a dramatic change between the two culturing conditions. We also detected micro-rheological differences in the cell periphery between the cells cultured in these two media correlating well with the observed nano-architecture of the ESCs in the two different culture conditions. These results pave the way for linking physical properties and cytoskeletal architecture to cell morphology during early development.

scholarly journals Changes in Cell Morphology and Actin Organization in Embryonic Stem Cells Cultured Under Different Conditions

2021 ◽  
Author(s):  
Kasper Graves Hvid ◽  
Irene Istúriz Petitjean ◽  
Younes Farhangi Barooji ◽  
Joshua M Brickman ◽  
Lene Broeng Oddershede ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Cell Morphology ◽  
Ex Vivo ◽  
Embryonic Stem ◽  
Super Resolution ◽  
Cell Periphery ◽  
Regulatory Structure ◽  
Actin Structure ◽  
Cells Cultured

The cellular cytoskeleton provides the cell with a mechanical rigidity which allows mechanical interaction between cells and the extracellular environment. The actin structure plays a key role in mechanical events like motility, or establishment of cell polarity. From the earliest stages of development, as represented by ex vivo expansion of naïve embryonic stem cells (ESCs), the critical mechanical role of the actin structure is becoming recognized as a vital cue for correct segregation and lineage control of cells and as a regulatory structure that controls several transcription factors. Naïve ESCs have a characteristic morphology and the ultrastructure that underlies this condition remains to be further investigated. Here, we investigate the 3D actin cytoskeleton of naïve mouse ESCs using super resolution optical reconstruction microscopy (STORM). We investigate the morphological, cytoskeletal and mechanical changes in cells cultured in 2i or Serum/LIF media reflecting a homogenous preimplantation cell state and a state that is closer to embarking on differentiation. STORM imaging showed that the peripheral actin structure undergoes a dramatic change between the two media conditions. We also detected micro-rheological differences in the cell periphery between the cells cultured in these two media correlating well with the observed nano-architecture of the ESCs in the two different culture conditions. These results pave the way for linking physical properties and cytoskeletal architecture to cell morphology during early development.

scholarly journals Physical properties and actin organization in embryonic stem cells depend on differentiation stage

2020 ◽  
Author(s):  
K. G. Hvid ◽  
Y. F. Barooji ◽  
I. Isturiz ◽  
J. M. Brickman ◽  
L.B. Oddershede ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Physical Properties ◽  
Embryonic Stem ◽  
Super Resolution ◽  
Stages Of Development ◽  
Actin Organization ◽  
Cytoskeletal Structure ◽  
Actin Structure ◽  
Differentiation Stage

AbstractThe cellular cytoskeleton provides the cell with mechanical rigidity and mediates mechanical interaction between cells and with the extracellular environment. The actin structure plays a key role in regulating cellular behaviors like motility, cell sorting, or cell polarity. From the earliest stages of development, in naïve stem cells, the critical mechanical role of the actin structure is becoming recognized as a vital cue for correct segregation and lineage control of cells and as a regulatory structure that controls several transcription factors. The ultrastructure of the earliest embryonic stem cells has not been investigated in living cells despite the fact that it is well-known that cells undergo morphological shape changes during the earliest stages of development. Here, we provide 3D investigations of the actin cytoskeleton of naïve mouse embryonic stem cells (ESCs) in clusters of sizes relevant for early stage development using super resolution optical reconstruction microscopy (STORM). We quantitatively describe the morphological, cytoskeletal and mechanical changes appearing between cells in small clusters at the earliest stages of inner cell mass differentiation, as recapitulated by cells cultured under two media conditions, 2i and Serum/LIF, thus promoting the naïve and first primed state, respectively. High resolution images of living stem cells showed that the peripheral actin structure undergoes a dramatic change between the two media conditions. The actin organization changed from being predominantly oriented parallel to the cell surface in 2i medium to a more radial orientation in Serum/LIF. Finally, using an optical trapping based technique, we detected micro-rheological differences in the cell periphery between the cells cultured in these two media, with results correlating well with the observed nano-architecture of the ESCs in the two different differentiation stages. These results pave the way for linking physical properties and cytoskeletal architecture to the development from naïve stem cells to specialized cells.Statement of SignificanceCells receive mechanical signals and must provide mechanical feedback, therefore, physical properties are instrumental for cell-cell interactions. Mechanical signals mediated through the cell surface can significantly affect transport of signaling molecules and can influence biological processes like transcriptional regulation. To achieve a deeper insight into how the cytoskeletal structure is responsible for cell shape and material properties at the earliest stages of development, we employ super-resolution microscopy to image actin fibers in clusters of embryonic stem cells mimicking early development. By modification of the culturing conditions, we investigate how the actin cytoskeleton and micro-rheological properties of ESCs change between the naïve ground state and the stage primed towards epiblast, thus revealing a correlation between differentiation stage and cytoskeletal structure.

scholarly journals LIF removal increases CRABPI and CRABPII transcripts in embryonic stem cells cultured in retinol or 4-oxoretinol

2008 ◽  
Vol 280 (1-2) ◽  
pp. 63-74 ◽  
Author(s):  
Michelle A. Lane ◽  
Juliana Xu ◽  
Elana W. Wilen ◽  
Renia Sylvester ◽  
Fadila Derguini ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Cells Cultured

106 STUDIES OF NUCLEAR ARCHITECTURE IN MAMMALIAN PRE-IMPLANTATION EMBRYOS AND EMBRYONIC STEM CELLS USING SUPER-RESOLUTION FLUORESCENCE MICROSCOPY

2013 ◽  
Vol 25 (1) ◽  
pp. 200
Author(s):  
J. Popken ◽  
M. Sterr ◽  
Y. Markaki ◽  
M. Cremer ◽  
A. Beck ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Fluorescence Microscopy ◽  
Nuclear Organization ◽  
Embryonic Stem ◽  
Super Resolution ◽  
Nuclear Bodies ◽  
Interchromatin Compartment ◽  
Cold Spring ◽  
Perichromatin Region

Three-dimensional (3-D) super-resolution fluorescence microscopy has allowed major progress in studies of the functional nuclear organization (Markaki et al. 2010 Cold Spring Harb. Symp. Quant. Biol. 75, 475–492; Markaki et al. 2012 Bioessays 34, 412–426). We have exploited these new possibilities to explore nuclear organization at different stages of bovine pre-implantation development (4-cell, 8-cell, 16-cell, morula, and blastocyst stage). In particular, we studied the topography of RNA polymerase II and the distribution of transcriptionally competent and noncompetent chromatin using antibodies against H3K4me3 and H3K27me3, respectively. For comparison, we have started analyses of mouse pre-implantation embryos and embryonic stem cells as well. Our results support the chromosome territory-interchromatin compartment (CT-IC) model (Cremer and Cremer 2010 Cold Spring Harb. Perspect. Biol. 2, a003889; Cremer et al. 2012 In: Epigenetic Regulation and Epigenomics 451–483). In all cell types, the nuclear space is occupied by chromosome territories (CTs; Koehler et al. 2009 Exp. Cell Res. 315, 2053–2063), the interchromatin compartment (IC), and one or several nucleoli. The CTs are built up from interconnected, megabase-sized chromatin domains (CDs). These ~1-Mbp CDs may consist of a series of ~100-kbp CDs (Cremer et al. 2000 Crit. Rev. Eukaryot. Gene Expr. 10, 179–212), which globally form a compact chromatin core surrounded by a layer of decondensed chromatin, called the perichromatin region. Current evidence supports the hypothesis that the perichromatin region represents the nuclear compartment, where transcription, co-transcriptional splicing, DNA-replication, and DNA-repair take place (Rouquette et al. 2010 Int. Rev. Cell Mol. Biol. 282, 1–90). The IC provides a contiguous, crowded compartment, which starts with channels at nuclear pores and pervades the chromatin compartment both between and within CTs. Small-scale chromatin loops of the perichromatin region can protrude into the interior of IC channels allowing direct contacts between CDs in cis and trans. At other sites the IC expands to wider, chromatin-free lacunas with splicing speckles and nuclear bodies. This model is in line with a fractal higher-order chromatin arrangement at all levels from CTs, chromosome arms and bands to ~1 Mbp CDs organized as fractal globules (Mirny 2011 Chromosome Res. 19, 37–51). This work is supported by the DFG (ZA 425/1-3, CR 59/29-2).

scholarly journals RUNX1a enhances hematopoietic lineage commitment from human embryonic stem cells and inducible pluripotent stem cells

Blood ◽  
2013 ◽  
Vol 121 (15) ◽  
pp. 2882-2890 ◽  
Author(s):  
Dan Ran ◽  
Wei-Jong Shia ◽  
Miao-Chia Lo ◽  
Jun-Bao Fan ◽  
David A. Knorr ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Pluripotent Stem Cells ◽  
Ex Vivo ◽  
Ectopic Expression ◽  
Embryonic Stem ◽  
Embryoid Bodies ◽  
Lineage Commitment ◽  
Hematopoietic Differentiation

Abstract Advancements in human pluripotent stem cell (hPSC) research have potential to revolutionize therapeutic transplantation. It has been demonstrated that transcription factors may play key roles in regulating maintenance, expansion, and differentiation of hPSCs. In addition to its regulatory functions in hematopoiesis and blood-related disorders, the transcription factor RUNX1 is also required for the formation of definitive blood stem cells. In this study, we demonstrated that expression of endogenous RUNX1a, an isoform of RUNX1, parallels with lineage commitment and hematopoietic emergence from hPSCs, including both human embryonic stem cells and inducible pluripotent stem cells. In a defined hematopoietic differentiation system, ectopic expression of RUNX1a facilitates emergence of hematopoietic progenitor cells (HPCs) and positively regulates expression of mesoderm and hematopoietic differentiation-related factors, including Brachyury, KDR, SCL, GATA2, and PU.1. HPCs derived from RUNX1a hPSCs show enhanced expansion ability, and the ex vivo–expanded cells are capable of differentiating into multiple lineages. Expression of RUNX1a in embryoid bodies (EBs) promotes definitive hematopoiesis that generates erythrocytes with β-globin production. Moreover, HPCs generated from RUNX1a EBs possess ≥9-week repopulation ability and show multilineage hematopoietic reconstitution in vivo. Together, our results suggest that RUNX1a facilitates the process of producing therapeutic HPCs from hPSCs.

scholarly journals Embryonic Stem Cells Cultured in Microfluidic Chambers Take Control of Their Fate by Producing Endogenous Signals Including LIF

Stem Cells ◽  
2016 ◽  
Vol 34 (6) ◽  
pp. 1501-1512 ◽  
Author(s):  
Joshua Guild ◽  
Amranul Haque ◽  
Pantea Gheibi ◽  
Yandong Gao ◽  
Kyung Jin Son ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Microfluidic Chambers ◽  
Cells Cultured

scholarly journals Impairment of germline transmission after blastocyst injection with murine embryonic stem cells cultured with mouse hepatitis virus and mouse minute virus

2008 ◽  
Vol 18 (1) ◽  
pp. 45-57 ◽  
Author(s):  
E. Mahabir ◽  
K. Reindl ◽  
J. Mysliwietz ◽  
J. Needham ◽  
D. Bulian ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Hepatitis Virus ◽  
Mouse Hepatitis Virus ◽  
Embryonic Stem ◽  
Germline Transmission ◽  
Murine Embryonic Stem Cells ◽  
Minute Virus ◽  
Cells Cultured

scholarly journals Epithelial–mesenchymal transition process in human embryonic stem cells cultured in feeder-free conditions

2006 ◽  
Vol 13 (1) ◽  
pp. 21-32 ◽  
Author(s):  
U. Ullmann ◽  
P. In’t Veld ◽  
C. Gilles ◽  
K. Sermon ◽  
M. De Rycke ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Epithelial Mesenchymal Transition ◽  
Embryonic Stem ◽  
Transition Process ◽  
Mesenchymal Transition ◽  
Cells Cultured

scholarly journals Unique Gene Expression Signature by Human Embryonic Stem Cells Cultured Under Serum-Free Conditions Correlates with Their Enhanced and Prolonged Growth in an Undifferentiated Stage

Stem Cells ◽  
2006 ◽  
Vol 24 (1) ◽  
pp. 151-167 ◽  
Author(s):  
Heli Skottman ◽  
Anne-Marie Strömberg ◽  
Eija Matilainen ◽  
Jose Inzunza ◽  
Outi Hovatta ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Gene Expression Signature ◽  
Serum Free ◽  
Unique Gene ◽  
Serum Free Conditions ◽  
Cells Cultured
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