Patent ID: 9371512
Filing Date: 2016-06-21
CPC Classification: C12N

Claim Text:
1. A method for the production of a permanent human amniocytic cell line comprising: a) transfecting primary human amniocytic cells with a nucleic acid molecule comprising a nucleic acid sequence encoding the human adenoviral serotype 5 gene products E1A and E1B to produce clonal transformed human amniocytic cells; b) isolating and expanding the clonal cells to produce a single transformed human amniocytic cell clone; c) expanding the single transformed human amniocytic cell clone of step (b) to produce a uniform transformed human amniocytic cell line having stable, high growth density and transient protein expression, and d) stably transfecting the transformed human amniocytic cells of step (c) with a nucleic acid molecule comprising a nucleic acid sequence encoding the SV40 large T-antigen operably linked to a CMV promoter or CAG promoter to produce a permanent human amniocytic cell line expressing SV40 large T-antigen, wherein said permanent human amniocytic cell line exhibits about a 25 to 70 fold increase in transient expression of recombinant protein compared to amniocytic cells not expressing SV40 large T-antigen when subsequently transfected with a nucleic acid encoding the recombinant protein.