In vitro generation of functional murine heart organoids via FGF4 and extracellular matrix
Lee, J., Sutani, A., Kaneko, R., Takeuchi, J., Sasano, T., Kohda, T., Ihara, K., Takahashi, K., Yamazoe, M., Morio, T., Furukawa, T., & Ishino, F.
Nature Communications (2020)
Mark Ghaly, Annabelle Nettey, and Tanyaradzwa Makunike
Background Information
Cardiogenesis
(Chong et al., 2014)
Embryonic Mouse Heart Development
(Lee et al., 2020)
(Sigma Aldrich, n.d.)
FGF Superfamily
(Sigma Aldrich, n.d.)
FGF Superfamily
FGF4
FGF10
Laminin-Entactin (LN/ET) Complex
(Arends & Lieleg, 2016)
Heart Organoid: the “Mini Heart”
(Betts et al., 2013; Hastings, 2021)
Research Goal
Generate a heart organoid similar to an embryonic mouse heart to determine key components of cardiogenesis
Hypothesis
The presence of fibroblast growth factor 4 (FGF4) and the laminin-entactin (LN/ET) complex is crucial for cardiogenic differentiation and heart formation
Methods
Isolating Embryonic Stem Cells
Formation of Embryonic Bodies
+ LIF
In vitro cardiogenesis, adding:
Experimental Design — LN/ET complex
LN/ET complex + FGF4
Matrigel
complex + FGF4
FGF4 only
(Control)
Cultured samples for 10-15 days
Analyzing morphological changes + heart organoid beat (bpm)
Isolating Embryonic Stem Cells
Formation of Embryonic Bodies
+ LIF
In vitro cardiogenesis; adding LN/ET complex and either:
Experimental Design — FGF Signals
FGF`10
FGF4
Cultured samples for 10-15 days
Analyzing heart organoid formation efficiency (%)
Results
Embryonic Murine Heart Development
Heart organoid development in embryonic bodies cultured in LN/ET complex + FGF4
Morphological changes and beating capacity were observed at day 3 and increased by day 6
Heart organoid growth in embryoid bodies cultured in Matrigel is constrained vs. LN/ET complex
Heart organoid generation is less
efficient in embryonic bodies cultured in
LN/ET + FGF10 compared to LN/ET + FGF4
Heart organoid generation is less
efficient in embryonic bodies cultured in
LN/ET + FGF10 compared to LN/ET + FGF4
Conclusion
LN/ET complex and FGF4 are KEY components of embryonic heart development!
In vitro cardiogenesis of embryonic bodies cultured in a FGF4 and LN/ET complex mimics in vivo cardiogenesis in mice.
i.e. the authors’ hypothesis was supported
Future Studies
References
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Arends, F. & Lieleg, O. (2016). Biophysical properties of the basal lamina: A highly selective extracellular matrix. In Francesco Travascio (Ed.),
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Betts, J.G., Young, K.A., Wise, J.A., Johnson, E., Poe, B., Kruse, D.H., Korol, O., Johnson, J.E., Womble, M., & DeSaix, P. (2013, April 25). An
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2021/05/cardiac-organoids-self-organize-to-mimic-human-heart.html
Sigma Aldrich. (n.d.). Fibroblast growth factor family (FGF). https://www.sigmaaldrich.com/CA/en/technical-documents/technical-article/
cell-culture-and-cell-culture-analysis/mammalian-cell-culture/fibroblast-growth
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