Post-partum mammalian placenta, its use and placental stem cells therefrom

Abstract

The present invention provides a method of extracting and recovering embryonic-like stem cells, including, but not limited to pluripotent or multipotent stem cells, from an exsanguinated human placenta. A placenta is treated to remove residual umbilical cord blood by perfusing an exsanguinated placenta, preferably with an anticoagulant solution, to flush out residual cells. The residual cells and perfusion liquid from the exsanguinated placenta are collected, and the embryonic-like stem cells are separated from the residual cells and perfusion liquid. The invention also provides a method of utilizing the isolated and perfused placenta as a bioreactor in which to propagate endogenous cells, including, but not limited to, embryonic-like stem cells. The invention also provides methods for propagation of exogenous cells in a placental bioreactor and collecting the propagated exogenous cells and bioactive molecules therefrom.

1 .- 59 . (canceled) 
     
     
         60 . A method of cryopreserving a cell population comprising placental cells, comprising: collecting a cell population from a post-partum placenta by passage of a perfusion fluid through either or both of the umbilical artery and umbilical vein of the placenta, wherein the collected cell population comprises placental cells, and cryopreserving the collected cell population. 
     
     
         61 . The method of  claim 60 , wherein platelets are removed from the cell population prior to cryopreservation. 
     
     
         62 . The method of  claim 60 , wherein de-nucleated cells are removed from the cell population prior to cryopreservation. 
     
     
         63 . The method of  claim 62 , wherein the de-nucleated cells are removed by centrifugation. 
     
     
         64 . The method of  claim 62 , wherein nucleated cells are isolated by density gradient centrifugation. 
     
     
         65 . The method of  claim 60 , wherein the perfusion fluid is an isotonic fluid comprising an anticoagulant. 
     
     
         66 . The method of  claim 65 , wherein the anticoagulant is heparin. 
     
     
         67 . A cryopreserved cell population produced by the method of  claim 60 . 
     
     
         68 . A cryopreserved cell population produced by the method of  claim 61 . 
     
     
         69 . A cryopreserved cell population produced by the method of  claim 62 . 
     
     
         70 . A cryopreserved cell population produced by the method of  claim 64 . 
     
     
         71 . The cryopreserved cell population of any of  claims 67 - 70 , wherein said cell population comprises CD34+ cells. 
     
     
         72 . The cryopreserved cell population of any of  claims 67 - 70 , wherein said cell population comprises CD34− cells. 
     
     
         73 . The cryopreserved cell population of any of  claims 67 - 70 , wherein said cell population comprises OCT-4+ and ABC-p+ cells.