Shape of 3D Culture
Initiative #3
Correlation of shape of Melanoma 3D Culture Masses with the expression of Metastasis-inducing genes
Background
Melanoma is a highly aggressive form of skin cancer that has an increasing rate of incidence worldwide. Most types of melanomas are aggressive and have already metastasized at the time of diagnosis. Traditional 2D cell culture models fail to mimic the complex three-dimensional architecture of tumors, limiting their predictive accuracy for in vivo behavior. Three-dimensional (3D) culture systems, such as spheroids or organoids, offer a more physiologically relevant environment to study tumor biology and metastasis. In 3D culture, the formed tumor mass shows different morphology, which correlates with the metastatic ability of the cancer. Less aggressive cancer cell lines show more spherical masses of well-defined borders. The more aggressive or metastatic cell lines form irregular masses of cells.
Hypothesis
We hypothesize that the shape of melanoma 3D culture masses will correlate with the expression of metastasis-inducing genes.
Objectives
Establish 3D cultures of different melanoma cell lines.
Evaluate and compare the colony shapes and morphologies formed by these cell lines in 3D culture.
Correlate colony morphology with known metastatic potentials of the melanoma cell lines.
Methods
1. Cell Lines
⦿ Select a panel of melanoma cell lines with known differences in metastatic potential.
Metastatic
Non or Low Metastatic
2. Establishment of 3D Cultures
⦿ Use Matrigel or collagen to establish 3D cultures (spheroids) for each melanoma cell line.
⦿ Seed cells in 96-well plates and optimize conditions for consistent colony formation (e.g., matrix concentration, cell density).
3. Colony Morphology Analysis
⦿ Use phase-contrast microscopy to image colonies over time
⦿ Analyze colony shape, size, and structural characteristics (e.g., roundness, surface texture).
⦿ Categorize colonies based on observed morphologies (e.g., compact, dispersed, invasive).
4. Correlation with Metastatic Potential
⦿ Compare the morphological categories with the known metastatic potential of each cell line.
⦿ Use statistical methods to determine the strength of the correlation.