Analysis of Cancer Stem Cell (CSC) by Flow Cytometry (CAT#: STEM-CBT-0076-WXH)

Introduction

In recent years, a special type of cancer cell—the cancer stem cell (CSC)—has been identified and characterized for different tumors. CSCs may be responsible for the recurrence of a tumor following a primarily successful therapy and are thought to bear a high metastatic potential.
The proportion of CSCs in tumor tissues is very small, only 0.01%-2.00%, which is difficult to isolate. Flow cytometry is helpful to seperate CSCs.
Multiparametric flow cytometry is the method of choice for the analysis of CSCs. It allows the simultaneous analysis of different cellular features with high performance and reliability. Moreover, it enables the separation of living cells on the basis of marker expression or functional properties by fluorescence-activated cell sorting. A major advantage of this technique is its ability to isolate rare cells, which is a prerequisite of identifying small cell populations within the tumor bulk. Quantification is also possible and can be achieved either by the addition of count check beads to the sample or by volume-based flow cytometry.

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Principle Applications Procedure Materials Notes Related Products

Principle

measurements of specific cluster of differentiation (CD) surface markers and stem cell-specific metabolic

Applications

• Cell Surface Marker Analysis
Similar to the CD of the hematopoietic lineages and other tissues , stem cells of solid tumors have been found to be identifiable by a set of cell surface markers. Different markers which have been successfully used in hematopoietic and mesenchymal stem cell analysis like CD34, CD133 (prominin-I), CD44, CD90 (Thy-1), CD105 (endoglin), and CD117 (c-kit) are also suitable for CSC characterization.
• Side Polulation Analysis
A unique characteristic of stem cells is their high expression of ABC transporter protein family members, which leads to the higher chemoresistance and drug resistance. This has been analyzed in studies with glioblastoma , colon carcinoma , breast cancer, and other cancer types. The high expression of ABC transporter proteins in CSCs enables a flow cytometric identification of these cells as a "side population".
• Sorting of CSCs
The proportion of CSCs in tumor tissues is very small, only 0.01%-2.00%, which is difficult to isolate. Flow cytometry is helpful to seperate CSCs.
• Determination of ALDH1 activity
ALDH1 is able to mediate resistance against chemotherapeutic drugs, its activity might be crucial for both stem cell longevity and the resistance of CSCs to chemotherapy.

Procedure

1.Sample acquisition
2.Sample clean-up
3.Dead cell exclusion
4.Sample staining
5.Multicolor analysis and quantification

Materials

CD34, CD133 (prominin-I), CD44, CD90 (Thy-1), CD105 (endoglin), and CD117 (c-kit)

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