Efficient definitive endoderm induction from mouse embryonic stem cell adherent cultures: A rapid screening model for differentiation studies
Definitive endoderm (DE) differentiation from mouse embryonic stem cell (mESC) monolayer cultures has been limited by poor cell survival or low efficiency. Recently, a combination of TGFβ and Wnt activation with BMP inhibition improved DE induction in embryoid bodies cultured in suspension. Based on...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2014-01-01
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| Series: | Stem Cell Research |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1873506113001542 |
| _version_ | 1819145108017119232 |
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| author | Josué Kunjom Mfopou Marloes Geeraerts Roba Dejene Stijn Van Langenhoven Asma Aberkane Leo A. Van Grunsven Luc Bouwens |
| author_facet | Josué Kunjom Mfopou Marloes Geeraerts Roba Dejene Stijn Van Langenhoven Asma Aberkane Leo A. Van Grunsven Luc Bouwens |
| author_sort | Josué Kunjom Mfopou |
| collection | DOAJ |
| description | Definitive endoderm (DE) differentiation from mouse embryonic stem cell (mESC) monolayer cultures has been limited by poor cell survival or low efficiency. Recently, a combination of TGFβ and Wnt activation with BMP inhibition improved DE induction in embryoid bodies cultured in suspension. Based on these observations we developed a protocol to efficiently induce DE cells in monolayer cultures of mESCs. We obtained a good cell yield with 54.92% DE induction as shown by Foxa2, Sox17, Cxcr4 and E-Cadherin expression. These DE-cells could be further differentiated into posterior foregut and pancreatic phenotypes using a culture protocol initially developed for human embryonic stem cell (hESC) differentiation. In addition, this mESC-derived DE gave rise to hepatocyte-like cells after exposure to BMP and FGF ligands. Our data therefore indicate a substantial improvement of monolayer DE induction from mESCs and support the concept that differentiation conditions for mESC-derived DE are similar to those for hESCs. As mESCs are easier to maintain and manipulate in culture compared to hESCs, and considering the shorter duration of embryonic development in the mouse, this method of efficient DE induction on monolayer will promote the development of new differentiation protocols to obtain DE-derivatives, like pancreatic beta-cells, for future use in cell replacement therapies. |
| first_indexed | 2024-12-22T12:52:47Z |
| format | Article |
| id | doaj.art-4fe5c98334b04bf6b1634174e69b2653 |
| institution | Directory Open Access Journal |
| issn | 1873-5061 1876-7753 |
| language | English |
| last_indexed | 2024-12-22T12:52:47Z |
| publishDate | 2014-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Stem Cell Research |
| spelling | doaj.art-4fe5c98334b04bf6b1634174e69b26532022-12-21T18:25:11ZengElsevierStem Cell Research1873-50611876-77532014-01-0112116617710.1016/j.scr.2013.10.004Efficient definitive endoderm induction from mouse embryonic stem cell adherent cultures: A rapid screening model for differentiation studiesJosué Kunjom Mfopou0Marloes Geeraerts1Roba Dejene2Stijn Van Langenhoven3Asma Aberkane4Leo A. Van Grunsven5Luc Bouwens6Cell Differentiation Unit, Diabetes Research Center, Vrije Universiteit Brussel, BelgiumCell Differentiation Unit, Diabetes Research Center, Vrije Universiteit Brussel, BelgiumCell Differentiation Unit, Diabetes Research Center, Vrije Universiteit Brussel, BelgiumCell Differentiation Unit, Diabetes Research Center, Vrije Universiteit Brussel, BelgiumCell Differentiation Unit, Diabetes Research Center, Vrije Universiteit Brussel, BelgiumLiver Cell Biology, Department of Cell Biology, Vrije Universiteit Brussel, BelgiumCell Differentiation Unit, Diabetes Research Center, Vrije Universiteit Brussel, BelgiumDefinitive endoderm (DE) differentiation from mouse embryonic stem cell (mESC) monolayer cultures has been limited by poor cell survival or low efficiency. Recently, a combination of TGFβ and Wnt activation with BMP inhibition improved DE induction in embryoid bodies cultured in suspension. Based on these observations we developed a protocol to efficiently induce DE cells in monolayer cultures of mESCs. We obtained a good cell yield with 54.92% DE induction as shown by Foxa2, Sox17, Cxcr4 and E-Cadherin expression. These DE-cells could be further differentiated into posterior foregut and pancreatic phenotypes using a culture protocol initially developed for human embryonic stem cell (hESC) differentiation. In addition, this mESC-derived DE gave rise to hepatocyte-like cells after exposure to BMP and FGF ligands. Our data therefore indicate a substantial improvement of monolayer DE induction from mESCs and support the concept that differentiation conditions for mESC-derived DE are similar to those for hESCs. As mESCs are easier to maintain and manipulate in culture compared to hESCs, and considering the shorter duration of embryonic development in the mouse, this method of efficient DE induction on monolayer will promote the development of new differentiation protocols to obtain DE-derivatives, like pancreatic beta-cells, for future use in cell replacement therapies.http://www.sciencedirect.com/science/article/pii/S1873506113001542 |
| spellingShingle | Josué Kunjom Mfopou Marloes Geeraerts Roba Dejene Stijn Van Langenhoven Asma Aberkane Leo A. Van Grunsven Luc Bouwens Efficient definitive endoderm induction from mouse embryonic stem cell adherent cultures: A rapid screening model for differentiation studies Stem Cell Research |
| title | Efficient definitive endoderm induction from mouse embryonic stem cell adherent cultures: A rapid screening model for differentiation studies |
| title_full | Efficient definitive endoderm induction from mouse embryonic stem cell adherent cultures: A rapid screening model for differentiation studies |
| title_fullStr | Efficient definitive endoderm induction from mouse embryonic stem cell adherent cultures: A rapid screening model for differentiation studies |
| title_full_unstemmed | Efficient definitive endoderm induction from mouse embryonic stem cell adherent cultures: A rapid screening model for differentiation studies |
| title_short | Efficient definitive endoderm induction from mouse embryonic stem cell adherent cultures: A rapid screening model for differentiation studies |
| title_sort | efficient definitive endoderm induction from mouse embryonic stem cell adherent cultures a rapid screening model for differentiation studies |
| url | http://www.sciencedirect.com/science/article/pii/S1873506113001542 |
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