BME Seminar Series: Siddarth Chandrasekaran, M.S. Candidate

In vitro Method for Enriching Cancer Stem Cell Subpopulations in Melanoma using PDMS

Advisor: Lisa DeLouise, Ph.D., M.P.D.

Abstract

Cancer Stem Cells (CSCs) are a subpopulation of cells within a tumor exhibiting increased in vivo tumorigenicity, capacity for indefinite potential for in vivo self-renewal and the ability to generate phenotypically diverse cells including non-tumorigenic cells present in the tumor. Isolation of CSCs and studying their properties hold a lot of therapeutic promise. In vitro spheroid cell culture has been proved to be an assay for identifying malignant melanoma initiating cells (i.e. CSC). We address the presence of stem-like cells in an established tumorigenic melanoma cell line WM115 by culturing them on a commonly used silicone based polymer polydimethylsiloxane (PDMS).

PDMS is an elastomeric hydrophobic substrate that hinders cell adhesion and cells assume a more 3D morphology. 3D configuration of cells is known to mimic the in vivo conditions. Through this work we have developed a novel in vitro spheroid cell culture method to rapidly enrich non-adherent sub-populations that express higher levels of stem cell markers using melanoma cell lines. We also intend to determine the clonogenic potential of isolated CSC sub-populations using microbubbles, small spherical cavities (200-300 µm in diameter) formed in PDMS. Future application of this technology will be to isolate CSC in primary and metastatic tumors.