Cell Growth, Apoptosis and 3D Growth
Many of our clients want deeper insight into the in vitro growth of their selected tumor system. The interplay between tumor biology and drug treatment can alter the balance between cell division, cell death and inter-cellular communication. This can be modeled in 2D and 3D cell cultures.
We offer tools to study:
– Cell growth in 2D
– Cell death, including apoptosis
– 3D growth and morphology
In 2D cultures, knockdown of retinoblastoma protein (RB) gene expression had no detectable effect on cell growth in 4 breast cancer cell lines (MCF-7ras, MCF-7, MCF10A, ZR-75) A short hairpin RNA (shRb-1) designed to block Rb expression was delivered by a lentiviral vector.
In 3D culture systems, knockdown of RB gene expression promoted formation of mammo-spheroid growth (bar chart, right) with an increase in protrusions (image panel, left) that was generated by invading cells and cell clusters (arrows).
MCF7ras and T47D were selected for their ability to form mammospheres in semi-solid cultures containing methylcellulose.
These 3D studies highlight the need to select appropriate model systems to uncover important biological phenomena that may otherwise be overlooked in 2D cell cultures.
Cell proliferation and other markers can be detected in tissue samples and subsequently quantified. An example here are B16F18 mouse melanoma cells growing as metastases in C57bl/6 female mice, whose proliferation is detected using antibody against Ki67 proliferation antigen.