Hello,
I am currently exploring different thymidine treatments on Hela Cells. However, my Fascan profiles consistently reveal two peaks, suggesting that none of the thymidine conditions are achieving successful synchronization. I suspect mycoplasma contamination. What are the optimal concentration and duration conditions currently recommended for successful synchronization in cell cultures? Additionally, how might mycoplasma contaminations impact treatments involving thymidine? Any suggested protocols or references would be greatly appreciated.
It seems like you're asking about two different topics: contamination in cell culture and thymidine treatment. Let's address each one:
Contamination in Cell Culture: Contamination in cell culture is a common issue that can compromise experimental results. Contaminants may include bacteria, fungi, mycoplasma, or other cell lines. To prevent contamination, it's crucial to maintain sterile conditions during cell culture handling, including working in a laminar flow hood, using sterile techniques, and regularly testing for contaminants. Routine microscopic examination, culture morphology assessment, and periodic testing for mycoplasma are common practices to detect contamination.
Thymidine Treatment: Thymidine is a nucleoside analog that can be used in cell culture experiments to synchronize cells at the G1/S boundary of the cell cycle. By adding thymidine to the culture medium, cells are temporarily blocked from progressing through the cell cycle. After a period of thymidine treatment, the compound is removed, allowing synchronized cells to progress synchronously through the cell cycle upon release.
Thymidine treatment can be useful for studying various aspects of cell cycle regulation, DNA replication, and related processes. However, it's essential to optimize thymidine concentration and treatment duration based on the specific cell type and experimental objectives, as excessive thymidine treatment can induce cellular stress responses or other unintended effects.
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