Comparison of Differentiation Pattern and WNT/SHH Signaling in Pluripotent Stem Cells Cultured under Different Conditions
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Culture
2.2. In Vitro Differentiation of ESCs
2.3. WNT11 Treatment
2.4. Modulation of WNT Signaling Pathways
2.5. SHH Treatment
2.6. RNA Isolation and qPCR Analysis
2.7. Flow Cytometry Analysis
2.8. Immunolocalization
2.9. Western Blotting
2.10. ELISA
2.11. Statistical Analysis
3. Results
3.1. ESCs Preferentially Differentiate into Ectodermal Lineage under Defined Culture Conditions
3.2. Wnt Expression and Activity Significantly Differ in Mouse ESCs Cultured in the Presence of SR or FCS
3.3. Both Canonical and Noncanonical WNT Signaling Pathway Are Crucial for ESC Differentiation into Mesoderm
3.4. Higher SHH Expression and Activity Corresponds with Preferential Ectodermal Differentiation of Mouse ESCs
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Świerczek-Lasek, B.; Dudka, D.; Bauer, D.; Czajkowski, T.; Ilach, K.; Streminska, W.; Kominek, A.; Piwocka, K.; Ciemerych, M.A.; Archacka, K. Comparison of Differentiation Pattern and WNT/SHH Signaling in Pluripotent Stem Cells Cultured under Different Conditions. Cells 2021, 10, 2743. https://0-doi-org.brum.beds.ac.uk/10.3390/cells10102743
Świerczek-Lasek B, Dudka D, Bauer D, Czajkowski T, Ilach K, Streminska W, Kominek A, Piwocka K, Ciemerych MA, Archacka K. Comparison of Differentiation Pattern and WNT/SHH Signaling in Pluripotent Stem Cells Cultured under Different Conditions. Cells. 2021; 10(10):2743. https://0-doi-org.brum.beds.ac.uk/10.3390/cells10102743
Chicago/Turabian StyleŚwierczek-Lasek, Barbara, Damian Dudka, Damian Bauer, Tomasz Czajkowski, Katarzyna Ilach, Władysława Streminska, Agata Kominek, Katarzyna Piwocka, Maria A. Ciemerych, and Karolina Archacka. 2021. "Comparison of Differentiation Pattern and WNT/SHH Signaling in Pluripotent Stem Cells Cultured under Different Conditions" Cells 10, no. 10: 2743. https://0-doi-org.brum.beds.ac.uk/10.3390/cells10102743