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Related Concept Videos

Cancer Stem Cells and Tumor Maintenance02:40

Cancer Stem Cells and Tumor Maintenance

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Early diagnosis and treatment can often cure cancer. However, even with treatment, residual cells called cancer stem cells (CSC) might remain, often causing tumor recurrence. These cancer stem cells possess the potential for self-renewal and multi-lineage differentiation and are often responsible for the therapeutic resistance displayed in most cancers.
Cancer stem cells are thought to originate from tissue-specific normal stem cells or progenitor cells. The normal stem cells usually reside in...
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Distinctive Features of Adult Stem Cells vs Cancer Stem Cells01:18

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A stem cell is an unspecialized cell that can divide without limit as needed and can, under specific conditions, differentiate into specialized cells.
Adult stem cells
Adult stem cells are tissue-specific; hence, they divide to develop the tissue from which they originate. One type of adult stem cell is the epithelial stem cell, which gives rise to the keratinocytes in the multiple layers of epithelial cells in the epidermis of the skin. Adult bone marrow has three distinct types of stem cells:...
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Stem Cell Culture01:17

Stem Cell Culture

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Stem cell research aims to find ways to use stem cells to regenerate and repair cellular damage. Over time, most adult cells undergo the wear and tear of aging and lose their ability to divide and repair themselves. Stem cells do not display a particular morphology or function. Adult stem cells, which exist as a small subset of cells in most tissues, keep dividing and can differentiate into a number of specialized cells generally formed by that tissue. These cells enable the body to renew and...
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Multipotency of Hematopoietic Stem Cells01:19

Multipotency of Hematopoietic Stem Cells

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The hematopoietic stem cells or HSCs are multipotent, meaning they can differentiate and give rise to all blood and immune cells. HSCs are maintained in the quiescent stage until an external stimulus initiates their differentiation. The multipotent HSCs exist as two heterogeneous populations, long-term repopulating cells (LTRC) and short-term repopulating cells (STRC). The two HSC populations have different surface markers or receptors and are classified based on quiescence and long-term...
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Related Experiment Video

Updated: Feb 21, 2026

Obtaining Cancer Stem Cell Spheres from Gynecological and Breast Cancer Tumors
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Self-Renewal and CSCs In Vitro Enrichment: Growth as Floating Spheres.

Pooja Mehta1, Caymen Novak2, Shreya Raghavan1

  • 1Department of Materials Science and Engineering, University of Michigan, Ann Arbor, MI, 48109-2800, USA.

Methods in Molecular Biology (Clifton, N.J.)
|October 8, 2017
PubMed
Summary

This chapter details methods for isolating cancer stem cells (CSCs) and culturing them in 3D spheroids using the hanging drop model. These techniques aid in understanding tumor biology and developing targeted cancer therapies.

Keywords:
Cancer stem cellsDrug sensitivityHanging dropOvarian cancerProliferationSpheroidViability

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Area of Science:

  • Cancer Biology
  • Stem Cell Research
  • Tumor Microenvironment

Background:

  • Cancer stem cells (CSCs) drive tumor progression and recurrence.
  • The 3D tumor microenvironment is crucial for CSC behavior.
  • 3D spheroids mimic the tumor microenvironment, serving as a valuable research tool.

Purpose of the Study:

  • To describe methodologies for CSC isolation and 3D spheroid culture.
  • To detail assays for characterizing CSCs within spheroids.
  • To provide guidance for successful 3D CSC spheroid experiments.

Main Methods:

  • Isolation of CSCs from tumors or cell lines using Fluorescence-Activated Cell Sorting (FACS).
  • Generation and maintenance of CSC spheroids using the 3D hanging drop model.
  • Assays for proliferation (alamarBlue) and viability (confocal microscopy, Live/Dead Kit), FACS characterization, and immunohistochemistry for cell interactions.

Main Results:

  • Established protocols for CSC isolation and 3D spheroid formation.
  • Quantifiable methods for assessing spheroid proliferation and viability.
  • Characterization of CSC populations and cell-matrix interactions within spheroids.

Conclusions:

  • The described methods enable robust study of CSCs in a 3D context.
  • These techniques are applicable across various cancer types for fundamental research.
  • The methodologies support drug screening and the development of preclinical chemotherapeutic strategies.