Decellularized placental membrane and methods of preparing and use thereof

Abstract

A method of preparing a decellularized placental membrane is provided. The method comprises removing cells from a pre-decellularized placental membrane comprising an amnion layer and a chorion layer to produce a decellularized placental membrane without separating the amnion layer from the chorion layer. The pre-decellularized placental membrane is obtained from an amniotic sac, and the decellularized placental membrane comprises the amnion layer and the chorion layer. Also provided is a decellularized placental membrane and a placenta-derived graft comprising the decellularized placental membrane. Further provided are the uses of the decellularized placental membrane or the placenta-derived graft.

1 - 27 . (canceled) 
     
     
         28 . A decellularized placental membrane comprising a decellularized amnion layer and a decellularized chorion layer in contact with each other, wherein the decellularized amnion layer is derived from an amnion layer of a pre-decellularized placental membrane after decellularization of the amnion layer in the pre-decellularized placental membrane, the decellularized chorion layer is derived from a chorion layer in the pre-decellularized placental membrane after decellularization of the chorion layer in the pre-decellularized placental membrane, and the amnion layer and the chorion layer remain in contact with each other during decellularization of the amnion layer and the chorion layer in the pre-decellularized placental membrane. 
     
     
         29 . The decellularized placental membrane of  claim 28 , wherein the decellularized placental membrane comprises one or more growth factors in an amount that is at least 10% greater than the sum of the amount of the one or more growth factors in a decellularized control isolated amniotic layer and the amount of the one or more growth factors in a decellularized control isolated chorionic layer each have a substantially identical area size as the decellularized placental membrane. 
     
     
         30 . The decellularized placental membrane of  claim 28 , wherein the decellularized placental membrane comprises one or more growth factors in an amount that is at least 20% of the amount of the one or more growth factors in the pre-decellularized placental membrane. 
     
     
         31 . The decellularized placental membrane of  claim 28 , wherein the decellularized placental membrane comprises platelet-derived growth factor (PDGF) in an amount that is at least 20% of the amount of the PDGF in the pre-decellularized placental membrane. 
     
     
         32 . The decellularized placental membrane of  claim 28 , wherein the decellularized placental membrane comprises basic fibroblast growth factor (bFGF) in an amount that is at least 20% of the amount of the bFGF in the pre-decellularized placental membrane. 
     
     
         33 . The decellularized placental membrane of  claim 28 , wherein the decellularized placental membrane comprises DNA in an amount that is less than 20% of the DNA in the pre-decellularized placental membrane. 
     
     
         34 - 35 . (canceled) 
     
     
         36 . The decellularized placental membrane of  claim 28 , wherein the decellularized placental membrane comprises less than 100 ng dsDNA per mg dry weight of the decellularized placental membrane. 
     
     
         37 . The decellularized placental membrane of  claim 28 , wherein the decellularized placental membrane comprises DNA in an amount that is less than 10% of the DNA in a pre-decellularized placental membrane. 
     
     
         38 . The decellularized placental membrane of  claim 28 , wherein the decellularized amnion layer comprises a fibroblast layer, and wherein the decellularized chorion layer comprises a reticular layer. 
     
     
         39 . The decellularized placental membrane of  claim 28 , wherein the decellularized amnion layer comprises epithelium, a basement membrane, a compact layer, a fibroblast layer, and a spongy layer, and wherein the decellularized chorion layer comprises a cellular layer, a reticular layer, a pseudo-basement membrane, and a trophoblast layer. 
     
     
         40 . The decellularized placental membrane of  claim 28 , wherein the decellularized amnion layer comprises a spongy layer. 
     
     
         41 . The decellularized placental membrane of  claim 28 , wherein the decellularized chorion layer comprises a trophoblast layer. 
     
     
         42 . The decellularized placental membrane of  claim 28 , wherein the decellularized amnion layer comprises a spongy layer and the decellularized chorion layer comprises a trophoblast layer. 
     
     
         43 . The decellularized placental membrane of  claim 28 , wherein the decellularized placental membrane is frozen or freeze-dried. 
     
     
         44 . The decellularized placental membrane of  claim 28 , wherein the decellularized placental membrane is sterilized. 
     
     
         45 . The decellularized placental membrane of  claim 28 , wherein the decellularized placental membrane is stored at room temperature or in cryopreservation. 
     
     
         46 . The decellularized placental membrane of  claim 45 , wherein the decellularized placental membrane has a water content of 0.01-5% by weight based on the total weight of the decellularized placental membrane. 
     
     
         47 . The decellularized placental membrane of  claim 45 , wherein the stored decellularized placental membrane has a water content of 5-95% by weight based on the total weight of the decellularized placental membrane. 
     
     
         48 . The decellularized placental membrane of  claim 28 , further comprising a water replacing agent. 
     
     
         49 . A placenta-derived graft comprising the decellularized placental membrane of  claim 28  and one or more agents.