Efficient non-viral transduction and culture of erythroid cells

Details Efficient non-viral transduction and culture of erythroid cells Efficient non-viral transduction and culture of erythroid cells

C - Chemistry, Metallurgy

12

N

C12N 15/18 (2006.01) C07K 14/71 (2006.01) C12N 5/06 (2006.01) C12N 5/08 (2006.01) C12N 5/10 (2006.01) C12N 15/85 (2006.01)

Patent

CA 2260332

The present invention involves 1) efficient non-viral gene transfer into human erythroid progenitors cells and 2) use of a constitutively active human EpoR gene to prolong the lifespan of progenitor cells. CD34' cells enriched from umbilical cord blood LDMNC are cultured er vivo at a low density in the absence of supportive stroma for 7 to days in a serum-free defined culture medium with the addition of IL-3, SCF and EPO. At this time, cells expand < 70 fold and hematopoietic progenitors are present at a frequency of 25 BFU-E, 28 CFU-E and 10 CFU-GM per 1 x 10 5 cells as detected by methylceliulose colony forming assay. Approximately 2% are CD34~ while 40% are GIyA~. Hematopoietic cells cultured in this manner are rendered highly susceptible to gene transfer by electroporation and a gene transfer effciency of >25% is obtained. The lifespan of erythroid progenitors that persist at the point of electroporation can be prolonged through the introduction of a constitutively active form of the human EpoR gene.This invention is useful for (1) genetic modification of hematopoietic stem cells, especially erythroid progenitors and (2) the propagation of erythroid cells in vitro for the production of hemoglobin.

Affiliated with

Also associated with

Rating

Efficient non-viral transduction and culture of erythroid cells does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Efficient non-viral transduction and culture of erythroid cells, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Efficient non-viral transduction and culture of erythroid cells will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFCA-PAI-O-2032522