Patent ID: 6048524
Filing Date: 2000-04-11
Classification: A61K,A61P,C07K,C12N

Abstract:
A method of expressing erythropoietin in a mammal, comprising the steps of:a) obtaining a source of primary cells from a mammal;b) transfecting primary cells obtained in (a) with a DNA construct comprising exogenous DNA encoding erythropoietin and additional DNA sequences sufficient for expression of the exogenous DNA in the primary cells, thereby producing transfected primary cells which express the exogenous DNA encoding erythropoietin;c) culturing a transfected primary cell produced in (b), which expresses the exogenous DNA encoding erythropoietin, under conditions appropriate for propagating the transfected primary cell which expresses the exogenous DNA encoding erythropoietin, thereby producing a clonal cell strain of transfected secondary cells from the transfected primary cell;d) culturing the clonal cell strain of transfected secondary cells produced in (c) under conditions appropriate for and sufficient time for the clonal cell strain of transfected secondary cells to undergo a sufficient number of doublings to provide a sufficient number of transfected secondary cells to produce erythropoietin; ande) introducing transfected secondary cells produced in (d) into a mammal of the same species as the mammal from which the primary cells were obtained in sufficient number to express erythropoietin in the mammal.