초록 ▼

A bioassay method for supporting human tumor stem cell colony growth is disclosed. The method is suitable for culture of a variety of neoplasms of differing histopathology. Tumor stem cell colonies arising from different types of cancer have differing growth characteristics and colony morphology. Th

A bioassay method for supporting human tumor stem cell colony growth is disclosed. The method is suitable for culture of a variety of neoplasms of differing histopathology. Tumor stem cell colonies arising from different types of cancer have differing growth characteristics and colony morphology. The present bioassay may be employed in clinical studies of the effects of anticancer drugs or irradiation on human tumor stem cells.

대표청구항 ▼

An in vitro method for quantitatively assaying for the viable colony-forming tumor stem cell content of a sepcimen of primary explanted cells obtained from primary or metastatic human tumors employing a two-layer culture system exhibiting interlayer diffusibility of dissolved nutrients and growth fa

An in vitro method for quantitatively assaying for the viable colony-forming tumor stem cell content of a sepcimen of primary explanted cells obtained from primary or metastatic human tumors employing a two-layer culture system exhibiting interlayer diffusibility of dissolved nutrients and growth factors, comprising the steps of: (a) forming a cell-free gelled underlayer comprising a liquid tissue culture feeder nutrient medium capable of supporting human tumor cell growth and a gelling agent for said feeder nutrient medium; (b) preparing a gelable liquid single-cell suspension of said explanted cells in a liquid tissue culture carrier nutrient medium capable of supporting human tumor cell growth and containing a gelling agent for said carrier nutrient medium; (c) plating said suspension onto said underlayer and allowing gelation thereof to occur, thereby forming a gelled overlayer which contains a known quantity of said explanted cells and which together with said underlayer constitutes said two-layer culture system, said culture system further containing a tumor stem cell colony growth-promoting concentration of METGF dissolved within at least one of said two layers, said METGF being a water-soluble tumor growth factor which is elaborated by macrophages; (d) incubating said culture system for a period of time sufficient to grow tumor stem cell colonies, the concentration of said explanted cells in said suspension being within a range enabling a substantially proportional relationship to exist between the total number of said explanted cells present in said overlayer and the total number of resulting tumor stem cell colonies grown during said incubation period; and (e) measuring the viable colony-forming tumor stem cell content of said overlayer as the total number of resulting tumor stem cell colonies grown during said incubation period.