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Updated: Jun 17, 2026
Analysis of Retinoic Acid-induced Neural Differentiation of Mouse Embryonic Stem Cells in Two and Three-dimensional Embryoid Bodies
Published on: April 22, 2017
Expression of a mutant retinoic acid receptor beta alters lineage differentiation in mouse embryonic stem cells
Christina Chatzi1, Christina E van den Brink, Paul T van der Saag
1School of Medical Sciences, Institute of Medical Sciences, University of Aberdeen , Foresterhill, Aberdeen, United Kingdom.
Abstract:
We have introduced 1 to 2 copies of a deletion mutant (betaDeltaC) of the human retinoic acid receptor beta into mouse embryonic stem (ES) cells. The betaDeltaC-expressing cells were 10 to 100 times less sensitive to RA-induced differentiation in comparison with their parental cells. In the presence of 10(-7) M RA in monolayer culture, they showed no growth arrest or differentiation, but remained pluripotent. Embryoid bodies (EBs) derived from betaDeltaC-expressing cells differentiated into cardiomyocytes rather than neurons after treatment with 10(-6) M RA, and became neurons upon exposure to 10(-5) or 10(-4) M RA. Remarkably, after 10 passages of continuous culture in the presence of 10(-7) M RA, they still were able to form chimeras after injection into blastocysts. These data suggest that appropriate levels of normal retinoid receptors are crucial for lineage-specific differentiation of mouse ES cells in vitro. The betaDeltaC mutant protein may prove to be useful in promoting "stemness" of ES cells in culture.
Insights
Introducing a retinoic acid receptor beta deletion mutant into mouse stem cells reduced differentiation sensitivity. These cells maintained pluripotency and chimera-forming ability, suggesting the mutant promotes stemness.
Area of Science:
- * Developmental Biology
- * Stem Cell Biology
- * Molecular Endocrinology
Background:
- * Retinoic acid (RA) signaling is critical for embryonic development and cell differentiation.
- * Retinoic acid receptor beta (RARβ) plays a key role in mediating RA's effects.
- * Mouse embryonic stem cells (ESCs) are a valuable model for studying early differentiation.
Purpose of the Study:
- * To investigate the role of retinoic acid receptor beta (RARβ) in ESC differentiation.
- * To determine if a deletion mutant of RARβ (RARβΔC) can affect ESC pluripotency and differentiation.
- * To explore the potential of RARβΔC in maintaining ESC stemness.
Main Methods:
- * Introduction of 1-2 copies of the human RARβΔC deletion mutant into mouse ESCs.
- * Assessment of RA-induced differentiation sensitivity in RARβΔC-expressing ESCs versus parental ESCs.
- * Culture of ESCs and embryoid bodies (EBs) under various RA concentrations.
- * Evaluation of pluripotency and chimera formation capacity after prolonged culture.
Main Results:
- * RARβΔC-expressing ESCs exhibited 10-100 fold reduced sensitivity to RA-induced differentiation.
- * In the presence of low RA concentrations (10⁻⁷ M), RARβΔC cells remained pluripotent without growth arrest or differentiation.
- * EBs derived from RARβΔC cells showed altered lineage commitment, differentiating into cardiomyocytes or neurons depending on RA concentration.
- * RARβΔC-expressing ESCs maintained chimera-forming ability after extensive passaging in RA.
Conclusions:
- * Specific levels of functional retinoid receptors are essential for lineage-specific differentiation of ESCs.
- * The RARβΔC mutant protein demonstrates potential for preserving ESC pluripotency and promoting stemness in culture.
- * Understanding RARβ function is crucial for controlling ESC differentiation and therapeutic applications.
