What is sphere forming assay?

What is sphere forming assay?

Sphere-formation assay is an in vitro method commonly used to identify CSCs and study their properties. Here, we report the detailed methodology on how to generate and propagate spheres from PCa cell lines and from murine prostate tissue.

How are cancer stem cells identified?

How are cancer stem cells identified? Researchers take samples of tumors removed from patients during surgery, always with the patient’s informed consent. The cells within the tumor are then sorted based on their expression of certain cell markers on their surface.

What is a sphere culture?

Sphere-forming culture was first used by Reynold and Weiss to isolate stem cells and has been widely used since [7]. In this technique, cells are attached to an extracellular matrix (ECM) that limits the growth, proliferation, and differentiation of the cells.

What is cancer stem cell theory?

The stem cell theory of cancer proposes that among all cancerous cells, a few act as stem cells that reproduce themselves and sustain the cancer, much like normal stem cells normally renew and sustain our organs and tissues.

How are spheres formed?

A sphere can also be constructed as the surface formed by rotating a circle about any of its diameters. Replacing the circle with an ellipse rotated about its major axis, the shape becomes a prolate spheroid; rotated about the minor axis, an oblate spheroid.

What is the origin of cancer stem cells?

Cancer stem cells might be derived from tissue-specific stem cells and bone marrow stem cells. They might also be derived from somatic cells that undergo transdifferentiation processes. Furthermore, cancer stem cells might be initiated as a result of cell fusion or horizontal gene-transfer processes.

What is the difference between cancer cells and cancer stem cells?

). For example, in tumors of the breast and brain, a minority population of cancer stem cells have the ability to self-renew, whereas the majority of cancer cells have limited or no ability to proliferate. This suggests that cancer stem cells may drive the growth and spread of the tumor.

What is the shape of sphere?

A sphere is a perfectly-round3D shapein the shape of a ball. All the points of its surface are equidistant (an equal distance) from its centre, meaning that it is smooth and has noedgesorvertices.

What are the three dimensions of a sphere?

Unlike a circle, which is a plane shape or flat shape, defined in XY plane, a sphere is defined in three dimensions, i.e. x-axis, y-axis and z-axis.

Is the sphere formation assay effective for cancer stem cells?

Although the existence of cancer stem cells (CSCs) has been demonstrated in colorectal cancer, further investigation is hindered by controversies over their surface markers. The sphere formation assay is widely used as in vitro method for derivation and characterization of CSCs based on the intrinsic self-renewal property of these cells.

How is sphere formation assay used in PCA?

In vitromodels to investigate the properties of CSCs in PCa are highly required. Sphere-formation assay is an in vitromethod commonly used to identify CSCs and study their properties.

Which is a reliable in vitro sphere formation assay?

Our results show the sphere-formation Assay (SFA) as a reliable in vitro assay to assess the presence and self-renewal ability of CSCs in different PCa models. This platform presents a useful tool to evaluate the effect of conventional or novel agents on the initiation and self-renewing properties of different tumors.

Can a colon sphere develop into a tumor?

Furthermore, cells that developed as colon spheres with strong self-renewal ability in vitro lost their tumorigenic capacity in vivo compared with Caco-2 cells, which could establish tumors in non-obese diabetic/severe-combined immunodeficient (NOD/SCID) mice.

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