IN VITRO TUMOR IN DISH KIT AND METHOD

• US 20130316392A1
• Filed: 05/20/2013
• Published: 11/28/2013
• Est. Priority Date: 05/24/2012
• Status: Active Grant

• Current Assignee: University of Kansas
• Original Assignee: University of Kansas
• Inventors: Anant, Shrikant, Ramamoorthy, Prabhu, Ramalingam, Satish, Sittampalam, Gurusingham Sitta
• Granted Patent: US 9,494,573 B2
• Time in Patent Office: Days
• US Class Current: 435/32
• CPC Class Codes: C12N 2502/1323 Adult fibroblasts
• CPC Class Codes: C12N 2502/28 Vascular endothelial cells
• CPC Class Codes: C12N 2503/02 Drug screening
• CPC Class Codes: C12N 5/0693 Tumour cells; Cancer cells
• CPC Class Codes: G01N 33/5011 for testing antineoplastic activity

Abstract

This invention is a method and kit for preparing a Tumor in Dish model for use in screening anti-cancer agents and analyzing cancer growth and development.

Claims

1. A kit for a Tumor in Dish model comprising culture medium or culture medium ingredients, isolated human umbilical vein endothelial cells, isolated human lymphatic endothelial cells, isolated human fibroblast cells, and isolated human epithelial cells.

2. The kit of claim 1, further comprising isolated cancer cells.

3. The kit of claim 1, further comprising an ultra low attachment culture vessel.

4. The kit of claim 1, wherein the culture medium or culture medium ingredients comprise Microvascular Endothelial Cell Growth Medium;

Airway Epithelial Basal Medium;

Dulbecco'"'"'s Modified Eagle'"'"'s Medium, Mammary Epithelial Cell Basal Medium, or Roswell Park Memorial Institute 1640 Medium;

Fibroblast Growth Medium;

Hydrocortisone;

Fibroblast growth factor 6;

Insulin-Like Growth Factor-I;

Ascorbic Acid;

Vascular Endothelial Growth Factor;

Human Epidermal Growth Factor;

Heparin;

Fetal Bovine Serum;

Antibiotics;

L-Alanyl-L-Glutamine;

Extract P;

Plasma Protein Fraction;

Epinephrine;

Transferrin;

Thyroxine 3;

Insulin; and

B27 supplement.

5. A method for preparing a Tumor in Dish model comprising

(a) combining isolated human umbilical vein endothelial cells, isolated human lymphatic endothelial cells, isolated human fibroblast cells, isolated human epithelial cells and isolated cancer cells;

and

(b) culturing the combination of cells in culture medium for a time sufficient to form a tumor thereby preparing a Tumor in Dish model.

6. The method of claim 5, wherein step (a) comprises

(i) combining isolated human umbilical vein endothelial cells, isolated human lymphatic endothelial cells, isolated human fibroblast cells and isolated human epithelial cells;

(ii) culturing the combination of cells of (i);

(iii) adding isolated cancer cells to the cultured cells of (ii).

7. The method of claim 5, wherein the step of culturing the cells is carried out in an ultra low attachment culture vessel.

8. The method of claim 5, wherein the culture medium comprises Microvascular Endothelial Cell Growth Medium;

Airway Epithelial Basal Medium;

Dulbecco'"'"'s Modified Eagle'"'"'s Medium, Mammary Epithelial Cell Basal Medium, or Roswell Park Memorial Institute 1640 Medium;

Fibroblast Growth Medium;

Hydrocortisone;

Fibroblast growth factor 6;

Insulin-Like Growth Factor-I;

Ascorbic Acid;

Vascular Endothelial Growth Factor;

Human Epidermal Growth Factor;

Heparin;

Fetal Bovine Serum;

Antibiotics;

L-Alanyl-L-Glutamine;

Extract P;

Plasma Protein Fraction;

Epinephrine;

Transferrin;

Thyroxine 3;

Insulin; and

B27 supplement.

9. The method of claim 5, further comprising steps:

(c) adding a test compound; and

(d) determining whether the test compound inhibits, attenuates, prevents or reduces tumor growth, proliferation, size or metastasis.