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.

What is claimed is:  
     
         1 . An isolated mammalian placenta which has been exsanguinated and perfused under sterile conditions.  
     
     
         2 . The isolated mammalian placenta of  claim 1  wherein the solution used to perfuse the placenta contains an anticoagulant solution.  
     
     
         3 . The isolated mammalian placenta of  claim 1  wherein the solution used to perfuse the placenta contains an antimicrobial solution.  
     
     
         4 . The isolated mammalian placenta of  claim 1  wherein the solution used to perfuse the placenta contains growth factors.  
     
     
         5 . The isolated mammalian placenta of  claim 1  wherein the placenta is human.  
     
     
         6 . The isolated mammalian placenta of  claim 1  which has been stored from about 2 to 24 hours after the expulsion of the placenta from the uterus.  
     
     
         7 . An isolated mammalian placenta which has been exsanguinated and perfused and incubated under conditions to allow for the production of embryonic-like stem cells and other multipotent stem cells from said placenta.  
     
     
         8 . The isolated mammalian placenta of  claim 7  which has been incubated for a period of about 2 to 24 hours.  
     
     
         9 . The isolated mammalian placenta of  claim 7  which has been perfused or incubated for a period of about 24 to more than 48 hours.  
     
     
         10 . An isolated, perfused mammalian placenta which comprises viable embryonic-like stem cells.  
     
     
         11 . The isolated placenta of  claim 10  wherein the stem cells are OCT-4− and ABC-p+.  
     
     
         12 . The isolated mammalian placenta of  claim 1  which has been perfused for at least two hours.  
     
     
         13 . The isolated mammalian placenta of  claim 12  which has been perfused for at least eleven hours.  
     
     
         14 . The mammalian placenta of  claim 10  wherein the placenta is recovered after birth.  
     
     
         15 . The isolated mammalian placenta of  claim 13  which has been perfused for at least twenty-four hours.  
     
     
         16 . The isolated mammalian placenta of  claim 10  wherein the placenta is perfused with a solution containing growth factors.  
     
     
         17 . The isolated mammalian placenta of  claim 7  which has been perfused with a solution containing growth factors.  
     
     
         18 . The isolated mammalian placenta of  claim 7  which is human.  
     
     
         19 . A method of culturing a mammalian placenta comprising obtaining a placenta after expulsion from the uterus, exsanguinating the placenta, and perfusing the placenta under sterile conditions.  
     
     
         20 . The method of  claim 19  wherein the placenta is perfused with a solution containing an anticoagulant solution.  
     
     
         21 . The method of  claim 19  wherein the placenta is perfused with a solution containing an antimicrobial agent.  
     
     
         22 . The method of  claim 19  wherein the placenta is human.  
     
     
         23 . The method of  claim 19  wherein the placenta has been stored for about two to twenty-four hours after expulsion from the uterus.  
     
     
         24 . The method of  claim 19  wherein said expulsion is at birth.  
     
     
         25 . The method of  claim 23  wherein the placenta is stored at room temperature.  
     
     
         26 . The method of  claim 23  wherein the placenta is stored under refrigeration or freezer conditions.  
     
     
         27 . A method for culturing an isolated mammalian placenta which has been exsanguinated and perfused comprising culturing the placenta under conditions to allow for the production of embryonic-like stem cells from said placenta.  
     
     
         28 . The method of  claim 27  wherein culturing the placenta comprises perfusing the placenta.  
     
     
         29 . The method of  claim 27  or  28  wherein the placenta has been incubated for a period of about twenty-four to forty-eight hours.  
     
     
         30 . The method of  claim 27  wherein the placenta has been perfused for at least two hours.  
     
     
         31 . The method of  claim 30  wherein the placenta has been perfused for at least eleven hours.  
     
     
         32 . The method of  claim 31  wherein the placenta has been perfused for at least twenty-four hours.  
     
     
         33 . The method of  claim 32  wherein the placenta has been preferred for more than forty-eight hours.  
     
     
         34 . The method of  claim 28  wherein the placenta has been perfused with a solution containing growth factors.  
     
     
         35 . The method of  claim 27  wherein the placenta is human.  
     
     
         36 . An isolated human placental stem cell which is OCT-4+ and ABC-p+.  
     
     
         37 . The stem of  claim 35  wherein the cell is a human cell.  
     
     
         38 . An isolated mammalian placental stem cell having at least the following characteristics: CD10+, CD29+, CD34−, CD44+, CD45−, CD54+, CD90+, SH2+, SH3+, SH4+, SSEA3−, SSEA4−, OCT-4+ and ABC-p+.  
     
     
         39 . The isolated placental stem cell of  claim 35  wherein said cell is SSEA3− and SSEA4−.  
     
     
         40 . A human placental stem cell which has been isolated from a post-partum human placenta after said placenta has been exsanguinated and perfused for at least 11 hours.  
     
     
         41 . A stem cell producing apparatus which comprises: 
 (a) a postpartum mammalian placenta which has been exsanguinated and perfused;    (b) a means for incubating or culturing the placenta; and    (c) a means for detecting stem cells.    
     
     
         42 . The apparatus of  claim 39  further comprising a collection device for collecting stem cells.  
     
     
         43 . The apparatus of  claim 39  further comprising a means for monitoring and adjusting the culture conditions.  
     
     
         44 . The apparatus of  claim 41  wherein the monitoring and adjusting is computerized.  
     
     
         45 . The apparatus of  claim 39  further comprising a cell separation device.  
     
     
         46 . A method of treating disease in a human which comprises administering a human placental stem of  claim 35 ,  37  or  38 .  
     
     
         47 . A method of transplanting stem cells which comprises administering to a patient in need thereof a human placental stem cell of  claim 35 ,  37  or  38 .  
     
     
         48 . A pharmaceutical composition which comprises a human placental stem cell of  claim 35 ,  37  or  38 .  
     
     
         49 . The pharmaceutical composition of  claim 46  which further comprises umbilical cord or placental blood.  
     
     
         50 . A committed cell which has been differentiated from a human placental stem cell of  claim 35 ,  37  or  38 .  
     
     
         51 . An isolated, homogenous population of human placental stem cells which can differentiate into all cell types.  
     
     
         52 . A homogenous population of viable human placental stem cells which exhibit at least the following cell surface markers: OCT-4+ and ABC-p+.  
     
     
         53 . An isolated, homogenous population of human placental stem cells which is multipotent.  
     
     
         54 . An isolated placenta containing a cell which is neither fetal nor maternal in origin.  
     
     
         55 . The stem cells of  claim 49  or  51  wherein the stem cells originate from a placenta.  
     
     
         56 . A composition suitable for bone marrow transplantation which comprises a population of hematopoietic stem cells enriched in cells that are CD34+ and CD38−.  
     
     
         57 . The composition of  claim 53  further comprising cord blood having cells that are CD34+ and CD38+.  
     
     
         58 . A composition suitable for bone marrow transplantation which comprises a population of hematopoietic stem cells enriched in cells that are CD34− and CD38−.  
     
     
         59 . The composition of  claim 54  further comprising cord blood having cells that are CD34+ and CD38+.