How to transfer a low metastasis cancer cell to a highly metastasis cancer cell in vitro?

Hello Dian Jamel

The metabolic differences among cancer cells confer differences in their metastatic potential. Metastatic cancer cells depend on monocarboxylate transporter 1 (MCT1) to deal with oxidative stress. MCT1 plays a major role in circulating lactate, which is a prominent energy source for metastasizing cells. As such, highly metastatic cells have increased levels of MCT1, whereas the inhibition of MCT1 decreases lactate uptake by metastatic cells and, thus, reduces their metastatic capability.

Also, changes in ATP/ADP and ATP/AMP ratios promote metastatic behavior.

So, having high levels of MCT1 on the surface of cells may probably help making the cells highly metastatic. Give it a thought.

Best.

Naseer Maliyakkal

Consider the cellular transformation by Oncogenic RAS and MMPs. Try Stepwise transfections.

Tomas Koltai

Dear Malcolm Nobre

I do not think that just by making a cell express METS the metastatic rate will increase. Metastasis is far more complex than that. On the other hand as Naseer says, transferring Ras can increase the number of metastasis. The simplest and cheapest way that can tested is to cultivate cells at a low oxygen level. Between 1 and1.5%. For a long time. Other option is concomitant culture in low oxygen environment with a pH under 6.5. Hypoxia plus acidity increase the metastatic phenotype.

Naseer Maliyakkal

But how will the cells grow under these conditions? Will it not die?

Saif Wahid

To generate highly metastatic lung cancer cell lines from low metastatic ones in vitro, you can employ a process called in vitro selection or serial passaging. Here's a general outline of the approach:

Culture Low Metastatic Cell Lines: Start by culturing your low metastatic lung cancer cell lines in appropriate cell culture conditions using standard protocols.

Prepare Appropriate Culture Conditions: Create an environment that mimics conditions that promote metastasis. This can include modifying the culture medium composition, adding specific growth factors, or altering oxygen levels.

Transwell Migration Assay: Use a Transwell migration assay to select and isolate cells with enhanced migratory capacity. This assay involves placing low metastatic cells in the upper chamber of a Transwell plate and providing a chemoattractant in the lower chamber. Cells that migrate through the pores to the lower chamber are collected and cultured separately.

Repeat Passaging: Take the migrated cells from the Transwell migration assay and expand their culture by passaging them in new culture dishes or flasks. Repeat this process for multiple rounds to increase the population of the selected cells.

Characterization: At each passage, characterize the cells for metastatic potential using various assays such as migration, invasion, or adhesion assays. This will help assess their progress towards acquiring a highly metastatic phenotype.

Validation: Confirm the metastatic properties of the generated cell lines by conducting in vitro and in vivo assays, such as wound healing assays, three-dimensional culture assays, or xenograft models in immunodeficient mice.

Genetic and Phenotypic Analysis: Perform genetic and phenotypic analyses, such as gene expression profiling, whole-genome sequencing, or proteomic analysis, to identify alterations that may contribute to the enhanced metastatic potential.

It's important to note that the metastatic potential of cell lines generated through in vitro selection may not fully recapitulate the complex processes observed in vivo. Therefore, it is crucial to validate the findings using relevant in vivo models and compare the results with patient-derived samples or established highly metastatic cell lines.

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