Selection of cells having increased cell adhesion properties
AssigneeLa Jolla Cancer Research Foundation
InventorsMichael D. Pierschbacher, Erkki I. Ruoslahti, Shoukat Dedhar
Life of this patent
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Abstract
A method for selecting cell lines expressing increased cell adhesion properties or promoting cell differentiation by culturing cells in increasing concentrations of adhesion ligand containing media. Cell lines produced by such method.
Claims20
A method for promoting differentiation of animal cells and preventing a differentiated state of animal cells from defifferentiating, comprising the steps of:
- a. providing an initial sample of said animal cells;
- b. culturing said animal cells in culture media containing an initial concentration of an RGD containing adhesion ligand in solution,
- wherein said RGD-containing adhesion ligand binds to RGD binding receptors on the surface of said animal cells which inhibits the adhesive function of said receptors, and wherein said initial concentration is selected so that not every receptor is bound;
- c. choosing those cells which proliferate in the presence of said initial concentration of said RGD containing adhesion ligand in solution;
- d. successively reculturing the chosen cells in culture media containing successively increasing concentrations of said adhesion ligand in solution;
- e. selecting those cells which proliferate in concentrations of said adhesion ligand in solution normally inhibitory to cell proliferation;
- and
- f. establishing from the selected cells a cell line which is more differentiated than said initial sample of animal cells.
A method for promoting the differentiation of animal cells and preventing a differentiated state of animal cells from dedifferentiating comprising the steps of:
- a. providing an initial sample of said animal cells;
- b. culturing said animal cells in culture media containing an initial concentration of an antibody in solution,
- wherein said antibody binds to RGD binding receptors on the surface of said animal cells which inhibits the adhesive function of said RGD binding receptors, and wherein said initial concentration is selected so that not every RGD binding receptor is bound;
- c. choosing those cells which proliferate in the presence of said initial concentration of said antibody in solution;
- d. successively reculturing said chosen cells in culture media containing successively increasing concentrations of said antibody in solution;
- e. selecting those cells which proliferate in concentrations of said antibody in solution normally inhibitory to cell proliferation;
- and
- f. establishing from said selected cells a cell line which is more differentiated than said initial sample of animal cells.
A method of increasing the differentiated state of undifferentiated malignant animal cells, normal undifferentiated animal stem cells, and dedifferentiated animal cells, comprising the steps of:
- a) reacting a soluble RGD-containing adhesion ligand at an initial concentration with a sample of animal cells selected from the group consisting of undifferentiated malignant animal cells, normal undifferentiated animal stem cells, and dedifferentiated animal cells;
- b) permitting said RGD-containing adhesion ligand to specifically bind to RGD binding cell surface receptors on the animal cells;
- c) identifying the subset of animal cells which have bound the highest concentration of said RGD-containing adhesion ligand to their RGD binding cell surface receptors;
- d) isolating culturing said subset of animal cells in the presence of a concentration of said RGD-containing adhesion ligand that is higher than the concentration in step (a);
- e) collecting the animal cells from step (d) and reacting them with said RGD-containing adhesion ligand at a concentration equal to the concentration of RGD-containing adhesion ligand in step (d);
- and
- f) repeating steps (b) through (e) until the differentiated state of the animal cells isolated in step (d) is increased.
A method for promoting the differentiation of animal cells and maintaining a differentiated state of animal cells likely to dedifferentiate, comprising the steps of:
- a) providing a soluble RGD-remaining adhesion ligand at an initial concentration wherein said RGD-containing ligand binds selectively to RGD binding cell surface receptors expressed by animal cells;
- b) permitting said RGD-containing ligand to react with a sample of animal cells;
- c) identifying the animal cells which have bound the highest concentration of said RGD-containing adhesion ligand to their RGD binding cell surface receptors;
- d) isolating and culturing said animal cells identified in step (c) in the presence of a concentration of said RGD-containing adhesion ligand that is higher than the concentration in step (a);
- e) collecting the animal cells from the culture of step (d) and reacting them with said RGD-containing adhesion ligand at a concentration equal to the concentration of RGD-containing adhesion ligand in step (d);
- and
- f) repeating steps (b) through (e) until the desired differentiated state of the animal cells isolated in step (d) is obtained.
Dependencies are inferred from the claim text.