Method for the separation of cells

摘要

The invention relates to a cell containing a gene encoding a conditional transgenic surface marker that is detectable upon expression on the surface of the cell, wherein the gene encoding a conditional transgenic surface marker comprises: (i) a promoter, operably linked to (ii) a first transcription sequence, and (iii) a second transcription sequence encoding the surface marker, whereby the first transcription sequence prevents the transcription of the second transcription sequence, whereby the first transcription sequence is conditionally removable such that the second transcription sequence is transcribable, and whereby the surface marker renders the cell sortable through the detection of the conditional transgenic surface marker. Furthermore, the invention relates to a construct for generating such a cell, and to a method for separating such a cell from a population of cells.

主权项

1. A cultured mammalian cell containing a gene encoding a conditionally expressed transgenic extracellular surface marker that is detectable on the surface of the cell and exhibits one or both of resistance to digestion with trypsin and non-internalization by the cell upon binding of an antibody, wherein the gene encoding a conditionally expressed transgenic surface marker comprises:
(i) a promoter for driving the transcription of a first transcription sequence, operably linked to (ii) a said first transcription sequence for preventing transcription of a second transcription sequence, thereby avoiding an immune response against the cell, wherein the first transcription sequence is conditionally removable by a CRE recombinase or a FLP recombinase, and (iii) a said second transcription sequence encoding the extracellular surface marker, comprising
(a) a transmembrane or membrane association domain that anchors the surface marker in the cell membrane wherein at least a portion of the surface marker is on the extracellular side of the cell membrane, and(b) a tag sequence for rendering a cell sortable, whereby the first transcription sequence is conditionally removable such that the second transcription sequence is transcribable, and whereby the surface marker renders the cell sortable through the detection of the conditionally expressed transgenic surface marker on the extracellular surface of the cell.