Isolation and Maintenance of Cancer Stem Cell - Creative Biolabs

Cancer stem cells (CSCs), also known as tumor-initiating cells, are a small population of cells within a tumor that have the ability to self-renew and differentiate into various cell types that constitute the tumor. The presence of CSCs in a tumor has important implications for cancer treatment. Researchers are actively studying CSCs to better understand their biology and develop new therapies that target these cells.

Creative Biolabs has been at the forefront of stem cell research and have developed a comprehensive protocol for the isolation and maintenance of CSCs. Our approach involves a meticulous step-by-step procedure that ensures the purity and viability of the stem cells. The protocol has been optimized to provide consistent and reliable results, and we have included detailed notes to help researchers troubleshoot any issues that may arise.

Materials Required

• Tissue culture dishes
• Medium
• Fetal bovine serum
• Penicillin-streptomycin
• Trypsin-EDTA
• Collagenase type IV
• Antibodies against stem cell markers

Procedure

Tissue Dissociation

The first step in the isolation of cancer stem cells is tissue dissociation. This involves breaking down the tissue into single cells using collagenase Type IV and trypsin-EDTA. The dissociated cells are then filtered to remove any debris and washed with HBSS.

FACS Sorting

The dissociated cells are stained with antibodies against stem cell markers and sorted using fluorescence-activated cell sorting (FACS).

Culture and Maintenance

The sorted cancer stem cells are then cultured in DMEM supplemented with FBS and P/S. The culture medium is changed every two days to ensure the viability of the stem cells. The stem cells are sub-cultured when they reach 70-80% confluency.

Notes

• The dissociation time and temperature should be optimized for each tissue type.
• The concentration of collagenase and trypsin-EDTA should be carefully titrated to avoid damaging the stem cells.
• The antibodies used for FACS sorting should be validated to ensure specificity.
• The culture medium should be changed every two days to maintain stem cell viability.

The isolation and maintenance of cancer stem cells is a complex process that requires a well-optimized protocol. Creative Biolabs has developed a comprehensive protocol that ensures the purity and viability of cancer stem cells. Our approach involves a step-by-step procedure, and we have included detailed notes to help researchers troubleshoot any issues that may arise.

With our protocol, researchers can confidently isolate and maintain cancer stem cells for a variety of applications, including disease modeling, drug discovery, and cell therapy. You can contact us directly to get more stem cell isolation services.

References

1. Majeti R, et al. Identification of a hierarchy of multipotent hematopoietic progenitors in human cord blood. Cell stem cell, 2007, 1(6): 635-645.
2. Valent P, et al. Cancer stem cell definitions and terminology: the devil is in the details. Nature Reviews Cancer, 2012, 12(11), 767-775.

For Research Use Only. Not For Clinical Use.