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A Quick and Efficient Method for the Purification of Endoderm Cells Generated from Human Embryonic Stem Cells
Published on: March 3, 2016
Klf4 and Klf5 differentially inhibit mesoderm and endoderm differentiation in embryonic stem cells
Irène Aksoy1, Vincent Giudice2, Edwige Delahaye3
11] Inserm, U846, 18 Avenue Doyen Lepine, Bron 69500, France [2] Stem Cell and Brain Research Institute, Bron 69500, France [3] Université de Lyon, Université Lyon 1, Lyon 69003, France [4] Genome Institute of Singapore, 60 Biopolis street, Singapore 138672, Singapore [5].
Abstract:
Krüppel-like factors (Klf) 4 and 5 are two closely related members of the Klf family, known to play key roles in cell cycle regulation, somatic cell reprogramming and pluripotency. Here we focus on the functional divergence between Klf4 and Klf5 in the inhibition of mouse embryonic stem (ES) cell differentiation. Using microarrays and chromatin immunoprecipitation coupled to ultra-high-throughput DNA sequencing, we show that Klf4 negatively regulates the expression of endodermal markers in the undifferentiated ES cells, including transcription factors involved in the commitment of pluripotent stem cells to endoderm differentiation. Knockdown of Klf4 enhances differentiation towards visceral and definitive endoderm. In contrast, Klf5 negatively regulates the expression of mesodermal markers, some of which control commitment to the mesoderm lineage, and knockdown of Klf5 specifically enhances differentiation towards mesoderm. We conclude that Klf4 and Klf5 differentially inhibit mesoderm and endoderm differentiation in murine ES cells.
Insights
Krüppel-like factor 4 (Klf4) and Krüppel-like factor 5 (Klf5) differentially regulate mouse embryonic stem cell differentiation. Klf4 inhibits endoderm formation, while Klf5 inhibits mesoderm formation.
Area of Science:
- Stem cell biology
- Developmental biology
- Gene regulation
Background:
- Krüppel-like factors (Klf) 4 and 5 are critical regulators of cell pluripotency and differentiation.
- Understanding their distinct roles is essential for controlling stem cell fate.
Purpose of the Study:
- To investigate the divergent functions of Klf4 and Klf5 in inhibiting mouse embryonic stem (ES) cell differentiation.
- To elucidate the specific lineages inhibited by each factor.
Main Methods:
- Gene expression analysis using microarrays.
- Chromatin immunoprecipitation coupled with high-throughput DNA sequencing (ChIP-seq).
- Gene knockdown experiments to assess differentiation outcomes.
Main Results:
- Klf4 negatively regulates endodermal markers and its knockdown promotes endoderm differentiation.
- Klf5 negatively regulates mesodermal markers and its knockdown promotes mesoderm differentiation.
- Klf4 and Klf5 exhibit distinct inhibitory roles on specific embryonic lineages.
Conclusions:
- Klf4 and Klf5 play opposing roles in directing mouse ES cell differentiation.
- Klf4 specifically inhibits endoderm differentiation, whereas Klf5 specifically inhibits mesoderm differentiation.
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