Ex Vivo Culture of Circulating Tumor Cells in the Cerebral Spinal Fluid from Melanoma Patients to Study Melanoma-Associated Leptomeningeal Disease

作者： Vincent Law1,2, Inna Smalley3, Brittany R. Evernden2, Margaret Baldwin4, Keiran S. M. Smalley1, Peter A. Forsyth1,2 1Department of Tumor Biology,University of South Florida, H. Lee Moffitt Cancer Center & Research Institute, 2Department of Neuro-Oncology,University of South Florida, H. Lee Moffitt Cancer Center & Research Institute, 3Department of Cancer Physiology,University of South Florida, H. Lee Moffitt Cancer Center & Research Institute, 4Department of Comparative Medicine,University of South Florida, H. Lee Moffitt Cancer Center & Research Institute

简介：

Overview

This article describes a protocol for the propagation of cerebral spinal fluid-circulating tumor cells (CSF-CTCs) collected from patients with melanoma-associated leptomeningeal disease (M-LMD) to develop preclinical models to study M-LMD.

Key Study Components

Area of Science

• Neuroscience
• Oncology
• Cell Biology

Background

• Leptomeningeal disease from melanoma (M-LMD) is a severe form of metastasis.
• CTCs infiltrate the cerebral spinal fluid, complicating treatment options.
• Understanding CTC biology is crucial for developing effective therapies.
• Current methodologies for studying CSF CTCs are limited.

Purpose of Study

• To propagate CSF CTCs from M-LMD patients.
• To generate preclinical models for studying M-LMD pathology.
• To facilitate molecular and functional analyses of M-LMD cells.

Methods Used

• Pre-coating T175 flasks with poly-l-lysine for cell culture.
• Using human meningeal cells to create conditioned media.
• Centrifugation and processing of cerebral spinal fluid samples.
• Establishing xenograft models in immunodeficient mice.

Main Results

• Successful propagation of CSF CTCs in vitro and in vivo.
• Insights gained into the CSF microenvironment's role in CTC survival.
• Identification of growth factors that enhance CTC culture.
• Development of a methodology for studying M-LMD effectively.

Conclusions

• Propagation of CSF CTCs provides a valuable tool for M-LMD research.
• Understanding the biology of CTCs can lead to new therapeutic strategies.
• Further refinement of methodologies is needed for optimal results.

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