Abstract
Cancer stem cells are considered the reservoir cancer cells that are resistant to most of the forms of cancer therapies and cause relapse of the tumor. Medulloblastoma (MB), a primary CNS tumor, is a very fast-growing tumor affecting younger population. In order to characterize medulloblastoma cancer stem cells or studying the drug resistance in MB mediated through the cancer stem cells, it becomes essential to isolate and study them. Isolation and characterization of tumor cells is a critical step in understanding the cancer progression and to devise novel approaches against them as drug targets. Typically, characterization of stem cells is done through surface marker analysis and with the advent of flow cytometry based techniques, this has become incredibly straightforward. Flow cytometry employs a uniformly linear flow of cells created by complex hydraulics of the flow cytometer followed by illuminating flow path with a LASER beam. This gives very valuable information about cell composition in forward scatter (FSC) and side scatter (SSC). The surface molecules of the cells can further be stained with various florescent dyes which upon excitation with the LASER beam will give the signal that will be detected by the instrument. Flow cytometer is high-throughput equipment and requires careful operation to get valuable information about the samples. In this chapter, we describe how from a bulk cell sample of medulloblastoma cells, cancer stem cells are isolated.
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Acknowledgements
The protocol described here and the data presented here are from Dr. Sheila Singh laboratory, McMaster University in Canada. The authors are thankful to her for providing the resources and giving the consent to publish.
Conflict of Interest
The authors declare no conflicts of interest.
Funding Statement
N.G. acknowledges the Science and Engineering Research Board grant, India (SB/S2/RJN-072/2015), (ECR/2017/00092) and the seed grant under Institute of Eminence Scheme (IOE), BHU for financial support. P.K. acknowledges Malviya postdoctoral fellowship under IOE Scheme, BHU.
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Kumar, P., Garg, N. (2022). Flow Cytometry Approaches to Obtain Medulloblastoma Stem Cells from Bulk Cultures. In: Dey, A., Malhotra, A., Garg, N. (eds) Medulloblastoma. Methods in Molecular Biology, vol 2423. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1952-0_8
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DOI: https://doi.org/10.1007/978-1-0716-1952-0_8
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