Two-stage culture protocol for isolating mesenchymal stem cells from amniotic fluid

Patent Number: 7101710

This patent useful for parties looking for methodologies to purify mesenchymal stem cells from amniotic fluid. The patent has 3 independent claims. Essentially all three are directed towards a two step culture process where amniotic fluid cells are first grown in a vessel, subsequently non-adherent cells are isolated, and allowed to expand. The difference between the claims is the media used and the manipulation procedures. It should be noted that all independent claims require the use of fetal calf serum, which may produce regulatory hurdles for clinical application of this technique. Amniotic fluid stem cells are of particular interest to the area of stem cell research since they express the pluripotency marker Oct-4 (Prusa AR et al Hum Reprod. 2003 Jul;18(7):1489-93), can differentiate into numerous tissues, and express similar markers to mesenchymal stem cells, that is: positive for CD90, CD105, CD166, CD49e, SH3, SH4, and HLA-ABC positive and negative for CD31, CD34, CD45, CD49d, CD123, and HLA-DR (In 't Anker PS et al. Stem Cells. 2004;22(7):1338-45.).

View this patent on the USPTO website.