Immunocompatible chorionic membrane products

Abstract

Provided herein is a placental product comprising an immunocompatible chorionic membrane. Such placental products can be cryopreserved and contain viable therapeutic cells after thawing. The placental product of the present invention is useful in treating a patient with a tissue injury (e.g. wound or burn) by applying the placental product to the injury. Similar application is useful with ligament and tendon repair and for engraftment procedures such as bone engraftment.

1 - 121 . (canceled) 
     
     
         122 . A placental product comprising a cryopreservation medium and a chorionic membrane, wherein:
 a. the chorionic membrane comprises viable therapeutic cells and is substantially free of trophoblasts, CD14+ macrophages, or both trophoblasts and CD14+ macrophage; and   b. the cryopreservation medium comprises a cryopreserving amount of a cryopreservative; and   c. optionally, the chorionic membrane is substantially free of vascularized tissue or vascularized tissue-derived immunogenic cells.   
     
     
         123 . The placental product of  claim 122 , wherein the viable therapeutic cells are native to the chorionic membrane and comprise MSCs, fibroblasts; stromal cells, or a combination thereof. 
     
     
         124 . The placental product of  claim 122 , wherein the cryopreservative comprises a cell-permeating cryopreservative, a non-cell-permeating cryopreservative, or a combination thereof, optionally wherein the cell-permeating cryopreservative comprises DMSO in amount of about 2% to about 20% 
     
     
         125 . The placental product of  claim 122 , wherein the viable therapeutic cells comprise stromal cells in an amount of about 5,000 to about 50,000 per cm 2  of the chorionic membrane. 
     
     
         126 . The placental product of any  claims 122 , wherein the chorionic membrane comprises fibroblasts in an amount of about 50% to about 90% of the total cells, wherein at least about 40% of the fibroblasts are viable after one freeze-thaw cycle, and wherein the fibroblasts are in an amount of at least about 7,000 cells/cm 2    
     
     
         127 . The placental product of  claim 122 , wherein the chorionic membrane comprises CD14+ macrophages in an amount of less than about 5%. 
     
     
         128 . The placental product of  claim 122 , wherein the chorionic membrane comprises functional macrophages in an amount of less than about 3,000 cells/cm 2 . 
     
     
         129 . The placental product of  claim 122 , wherein the chorionic membrane is substantially free of maternal dendritic cells, maternal leukocytes, or immunogenic maternal cells. 
     
     
         130 . The placental product of  claim 122 , comprising about 5,000 to about 50,000 stromal cells per cm 2 , wherein the viability of said stromal cells after one freeze-thaw cycle is about 70% to about 95% relative to fresh placental or chorionic membrane. 
     
     
         131 . The placental product of  claim 122 , wherein after one freeze-thaw cycle, the placental product or chorionic membrane releases TNF-α in an amount up to about 420 pg/mL into a tissue culture medium upon placing a 2 cm×2 cm piece of the placental product in a tissue culture medium and exposing the piece to a bacterial lipopolysaccharide for about 20 to about 24 hours. 
     
     
         132 . The placental product of  claim 122 , wherein the chorionic membrane further comprises one or more native factors selected from the group consisting of IGFBP1, adiponectin, α2-macroglobulin, bFGF, EGF, MMP-9 and TIMP1, wherein the one or more native factors are present in an amount, wherein the amount is substantially greater per gram of chorionic membrane than that of a fresh unprocessed chorionic membrane. 
     
     
         133 . The placental product of  claim 122 , further comprising an amniotic membrane wherein the amniotic membrane comprises a layer of amniotic epithelial cells. 
     
     
         134 . The placental product of  claim 122 , wherein the chorionic membrane comprises a basement membrane, a reticular layer, or a combination thereof. 
     
     
         135 . The placental product of  claim 122 , wherein the chorionic membrane has a thickness of about 40 μm to about 400 μm. 
     
     
         136 . The placental product of  claim 122 , wherein the maternal side of the chorionic membrane comprises fragments of extracellular matrix proteins in a concentration substantially greater than that of a native, unprocessed chorion. 
     
     
         137 . The placental product of  claim 122 , wherein the chorionic membrane has been treated with a protease, optionally wherein the protease is a dispase. 
     
     
         138 . The placental product of  claim 122 , wherein the chorionic membrane comprises a basement membrane, a reticular layer, or a combination thereof. 
     
     
         139 . The placental product of  claim 122 , wherein the chorionic membrane comprises MSCs, wherein:
 a. the MSCs are present in an amount of about 5% to about 30% relative to the total number of cells in the chorionic membrane;   b. at least about 40% of the MSCs are viable after one freeze-thaw cycle; and   c. the MSCs are present in an amount of at least about 1,500 cells/cm 2 .   
     
     
         140 . The method of  claim 139 , wherein:
 a. the viable therapeutic cells are native to the chorionic membrane and comprise MSCs;   b. the cryopreservative comprises DMSO in amount of about 2% to about 20%;   c. the viable therapeutic cells comprise stromal cells in an amount of about 5,000 to about 50,000 per cm 2  of the chorionic membrane, wherein the viability of said stromal cells after one freeze-thaw cycle is about 70% to about 95% relative to fresh placental or chorionic membrane;   d. the viable therapeutic cells comprise fibroblasts in an amount of about 50% to about 90% of the total cells, wherein the viability of the fibroblasts is at least about 40% after one freeze-thaw cycle, and wherein the fibroblasts are in an amount of at least about 7,000 cells/cm 2 ;   e. the chorionic membrane comprises CD14+ macrophages in an amount of less than about 5% of the total viable cells and less than about 3,000 cells/cm 2 ;   f. the chorionic membrane is substantially free of maternal dendritic cells, maternal leukocytes, and immunogenic maternal cells;   g. after one freeze-thaw cycle, the placental product or chorionic membrane release TNF-α up to about 420 pg/mL into a tissue culture medium upon placing a 2 cm×2 cm piece of the placental product in the tissue culture medium and exposing the piece to a bacterial lipopolysaccharide for about 20 to about 24 hours;   h. the chorionic membrane further comprises one or more native factors selected from the group consisting of IGFBP1, adiponectin, α2-macroglobulin, bFGF, EGF, MMP-9 and TIMP1; and   i. the chorionic membrane has a thickness of about 40 μm to about 400 μm.   
     
     
         141 . A method of treating a wound of a patient comprising administering the placental product of  claim 122  to the wound.