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08:21
In Vitro Differentiation of Human Pluripotent Stem Cells into Trophoblastic Cells
Published on: March 16, 2017
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Harnessing Autophagic Network Is Essential for Trophoblast Stem Cell Differentiation
Shreeta Chakraborty1, Rumela Bose1, Safirul Islam1
1Division of Cell Biology and Physiology, CSIR-Indian Institute of Chemical Biology, Kolkata, India.
Stem Cells and Development
|March 8, 2020
Summary
Autophagy, a cellular process, is upregulated during trophoblast stem cell differentiation. Inhibiting autophagy impairs placental cell development and reduces key placental cell markers, highlighting its crucial role.
Area of Science:
- Developmental Biology
- Cellular Biology
- Mammalian Placental Development
Background:
- Placental development involves trophoblast stem (TS) cell differentiation into specialized subtypes.
- This differentiation process requires dynamic proteome changes for specialized functions.
- Autophagy, a cellular degradation pathway, plays a role in cellular homeostasis.
Purpose of the Study:
- To investigate the role of autophagic machinery during trophoblast stem cell differentiation.
- To determine if autophagy is essential for the commitment and development of placental cell lineages.
- To elucidate the impact of autophagy inhibition on trophoblast cell differentiation markers and morphology.
Main Methods:
- Analysis of autophagic protein and LC3 maturation during TS cell differentiation.
- Assessment of Beclin1, Vps34, and PIK3R4 complex formation.
- Evaluation of p62/SQSTM1 degradation and LC3 colocalization in differentiating cells.
- Disruption of autophagy via lysosomal fusion inhibition in TS cells prior to differentiation induction.
- Quantification of placental cell differentiation markers (Prl3d1, Prl2c2, Prl4a1, Tpbpα) and assessment of trophoblast giant cell nuclear size.
Main Results:
- Autophagic machinery, including initiation and maturation proteins, is significantly upregulated during TS cell differentiation.
- Autophagic flux is triggered by differentiation induction, evidenced by LC3 maturation and enhanced Beclin1-Vps34-PIK3R4 complex formation.
- p62/SQSTM1 degradation and LC3 colocalization are prominent in trophoblast giant cells (TGCs) at day 6.
- Autophagy disruption leads to decreased expression of giant cell and spongiotrophoblast markers (Prl3d1, Prl2c2, Prl4a1, Tpbpα).
- Inhibition of autophagy results in reduced nuclear size in TGCs.
Conclusions:
- Autophagy is upregulated and actively triggered during trophoblast stem cell differentiation.
- Autophagy is essential for the proper differentiation of TS cells into specific placental lineages, including giant cells and spongiotrophoblasts.
- The study highlights autophagy's role in regulating protein turnover during the initial stages of trophoblast differentiation.
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