Simulated Microgravity Modulates Differentiation Processes of Embryonic Stem Cells
Background/Aims: Embryonic developmental studies under microgravity conditions in space are very limited. To study the effects of altered gravity on the embryonic development processes we established an in vitro methodology allowing differentiation of mouse embryonic stem cells (mESCs) under simulat...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Cell Physiol Biochem Press GmbH & Co KG
2016-04-01
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| Series: | Cellular Physiology and Biochemistry |
| Subjects: | |
| Online Access: | http://www.karger.com/Article/FullText/443090 |
| _version_ | 1818211596298616832 |
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| author | Vaibhav Shinde Sonja Brungs Margit Henry Lucia Wegener Harshal Nemade Tamara Rotshteyn Aviseka Acharya Christa Baumstark-Khan Christine E. Hellweg Jürgen Hescheler Ruth Hemmersbach Agapios Sachinidis |
| author_facet | Vaibhav Shinde Sonja Brungs Margit Henry Lucia Wegener Harshal Nemade Tamara Rotshteyn Aviseka Acharya Christa Baumstark-Khan Christine E. Hellweg Jürgen Hescheler Ruth Hemmersbach Agapios Sachinidis |
| author_sort | Vaibhav Shinde |
| collection | DOAJ |
| description | Background/Aims: Embryonic developmental studies under microgravity conditions in space are very limited. To study the effects of altered gravity on the embryonic development processes we established an in vitro methodology allowing differentiation of mouse embryonic stem cells (mESCs) under simulated microgravity within a fast-rotating clinostat (clinorotation) and capture of microarray-based gene signatures. Methods: The differentiating mESCs were cultured in a 2D pipette clinostat. The microarray and bioinformatics tools were used to capture genes that are deregulated by simulated microgravity and their impact on developmental biological processes. Results: The data analysis demonstrated that differentiation of mESCs in pipettes for 3 days resultet to early germ layer differentiation and then to the different somatic cell types after further 7 days of differentiation in the Petri dishes. Clinorotation influences differentiation as well as non-differentiation related biological processes like cytoskeleton related 19 genes were modulated. Notably, simulated microgravity deregulated genes Cyr61, Thbs1, Parva, Dhrs3, Jun, Tpm1, Fzd2 and Dll1 are involved in heart morphogenesis as an acute response on day 3. If the stem cells were further cultivated under normal gravity conditions (1 g) after clinorotation, the expression of cardiomyocytes specific genes such as Tnnt2, Rbp4, Tnni1, Csrp3, Nppb and Mybpc3 on day 10 was inhibited. This correlated well with a decreasing beating activity of the 10-days old embryoid bodies (EBs). Finally, we captured Gadd45g, Jun, Thbs1, Cyr61and Dll1 genes whose expressions were modulated by simulated microgravity and by real microgravity in various reported studies. Simulated microgravity also deregulated genes belonging to the MAP kinase and focal dhesion signal transduction pathways. Conclusion: One of the most prominent biological processes affected by simulated microgravity was the process of cardiomyogenesis. The most significant simulated microgravity-affected genes, signal transduction pathways, and biological processes which are relevant for mESCs differentiation have been identified and discussed below. |
| first_indexed | 2024-12-12T05:35:01Z |
| format | Article |
| id | doaj.art-788e6dce1cab4f66912f0f4e8ee3194e |
| institution | Directory Open Access Journal |
| issn | 1015-8987 1421-9778 |
| language | English |
| last_indexed | 2024-12-12T05:35:01Z |
| publishDate | 2016-04-01 |
| publisher | Cell Physiol Biochem Press GmbH & Co KG |
| record_format | Article |
| series | Cellular Physiology and Biochemistry |
| spelling | doaj.art-788e6dce1cab4f66912f0f4e8ee3194e2022-12-22T00:36:10ZengCell Physiol Biochem Press GmbH & Co KGCellular Physiology and Biochemistry1015-89871421-97782016-04-013841483149910.1159/000443090443090Simulated Microgravity Modulates Differentiation Processes of Embryonic Stem CellsVaibhav ShindeSonja BrungsMargit HenryLucia WegenerHarshal NemadeTamara RotshteynAviseka AcharyaChrista Baumstark-KhanChristine E. HellwegJürgen HeschelerRuth HemmersbachAgapios SachinidisBackground/Aims: Embryonic developmental studies under microgravity conditions in space are very limited. To study the effects of altered gravity on the embryonic development processes we established an in vitro methodology allowing differentiation of mouse embryonic stem cells (mESCs) under simulated microgravity within a fast-rotating clinostat (clinorotation) and capture of microarray-based gene signatures. Methods: The differentiating mESCs were cultured in a 2D pipette clinostat. The microarray and bioinformatics tools were used to capture genes that are deregulated by simulated microgravity and their impact on developmental biological processes. Results: The data analysis demonstrated that differentiation of mESCs in pipettes for 3 days resultet to early germ layer differentiation and then to the different somatic cell types after further 7 days of differentiation in the Petri dishes. Clinorotation influences differentiation as well as non-differentiation related biological processes like cytoskeleton related 19 genes were modulated. Notably, simulated microgravity deregulated genes Cyr61, Thbs1, Parva, Dhrs3, Jun, Tpm1, Fzd2 and Dll1 are involved in heart morphogenesis as an acute response on day 3. If the stem cells were further cultivated under normal gravity conditions (1 g) after clinorotation, the expression of cardiomyocytes specific genes such as Tnnt2, Rbp4, Tnni1, Csrp3, Nppb and Mybpc3 on day 10 was inhibited. This correlated well with a decreasing beating activity of the 10-days old embryoid bodies (EBs). Finally, we captured Gadd45g, Jun, Thbs1, Cyr61and Dll1 genes whose expressions were modulated by simulated microgravity and by real microgravity in various reported studies. Simulated microgravity also deregulated genes belonging to the MAP kinase and focal dhesion signal transduction pathways. Conclusion: One of the most prominent biological processes affected by simulated microgravity was the process of cardiomyogenesis. The most significant simulated microgravity-affected genes, signal transduction pathways, and biological processes which are relevant for mESCs differentiation have been identified and discussed below.http://www.karger.com/Article/FullText/443090Embryonic stem cellsDifferentiationTranscriptomicsSignal transduction pathwaysClinostatCardiomyogenesisMicrogravity |
| spellingShingle | Vaibhav Shinde Sonja Brungs Margit Henry Lucia Wegener Harshal Nemade Tamara Rotshteyn Aviseka Acharya Christa Baumstark-Khan Christine E. Hellweg Jürgen Hescheler Ruth Hemmersbach Agapios Sachinidis Simulated Microgravity Modulates Differentiation Processes of Embryonic Stem Cells Cellular Physiology and Biochemistry Embryonic stem cells Differentiation Transcriptomics Signal transduction pathways Clinostat Cardiomyogenesis Microgravity |
| title | Simulated Microgravity Modulates Differentiation Processes of Embryonic Stem Cells |
| title_full | Simulated Microgravity Modulates Differentiation Processes of Embryonic Stem Cells |
| title_fullStr | Simulated Microgravity Modulates Differentiation Processes of Embryonic Stem Cells |
| title_full_unstemmed | Simulated Microgravity Modulates Differentiation Processes of Embryonic Stem Cells |
| title_short | Simulated Microgravity Modulates Differentiation Processes of Embryonic Stem Cells |
| title_sort | simulated microgravity modulates differentiation processes of embryonic stem cells |
| topic | Embryonic stem cells Differentiation Transcriptomics Signal transduction pathways Clinostat Cardiomyogenesis Microgravity |
| url | http://www.karger.com/Article/FullText/443090 |
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