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Derivation of Mouse Trophoblast Stem Cells from Blastocysts
Published on: June 8, 2010
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Protocol for mouse trophoblast stem cell isolation, differentiation, and cytokine detection
Vijay Pratap Singh1, Jennifer L Gerton1,2
1Stowers Institute for Medical Research, 1000 E. 50th St, Kansas City, MO 64110, USA.
STAR Protocols
|January 18, 2021
Summary
This study details a protocol for isolating and culturing trophoblast stem cells. It also outlines methods for inducing genotoxic stress and analyzing the resulting changes in cytokine secretion, crucial for understanding pregnancy health.
Area of Science:
- Developmental Biology
- Cell Biology
- Immunology
Background:
- Trophoblast stem cells are vital for mammalian development, secreting factors essential for pregnancy.
- Pathological conditions can lead trophoblast cells to secrete proinflammatory cytokines, impacting maternal and fetal health.
Purpose of the Study:
- To provide a detailed protocol for trophoblast stem (TS) cell isolation, maintenance, and differentiation.
- To establish conditions for inducing genotoxic stress in differentiated TS cells.
- To detect the effects of genotoxic stress on cytokine production by TS cells.
Main Methods:
- Isolation and culture of trophoblast stem (TS) cells.
- Induction of genotoxic stress in differentiated TS cells.
- Detection and analysis of secreted factors, including cytokines.
Main Results:
- The protocol enables the study of TS cell behavior under stress conditions.
- Genotoxic stress was successfully induced in differentiated TS cells.
- The protocol facilitates the detection of changes in cytokine secretion.
Conclusions:
- This protocol offers a comprehensive method for studying trophoblast stem cells.
- It allows for the investigation of how genotoxic stress influences cytokine production in these critical cells.
- This research is valuable for understanding pregnancy pathologies.
