Immunocompatible amniotic membrane products

Abstract

Provided herein is a placental product comprising an immunocompatible amniotic 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 . A placental product comprising: a amniotic membrane, wherein the amniotic membrane:
 a. comprises viable therapeutic cells; and   b. is substantially free of functional CD14+ macrophages; and   optionally, wherein the viable therapeutic cells are native to the amniotic membrane.   
     
     
         2 . The placental product of  claim 1 , wherein the amniotic membrane is substantially free of vascularized tissue or vascularized tissue-derived immunogenic cells. 
     
     
         3 . The placental product of  claim 1 , wherein the amniotic membrane comprises a layer of epithelial cells. 
     
     
         4 . The placental product of  claim 1 , wherein the amniotic membrane comprises one, two, or three layers selected from: a compact layer, a layer of fibroblast, and a spongy layer. 
     
     
         5 . The placental product of  claim 1 , wherein the amniotic membrane is substantially free of immunogenic maternal cells. 
     
     
         6 . The placental product of  claim 1 , wherein the amniotic membrane is substantially free of maternal dendritic cells. 
     
     
         7 . The placental product of  claim 1 , wherein the amniotic membrane is substantially free of maternal leukocytes. 
     
     
         8 . The placental product of  claim 1 , wherein the viable therapeutic cells comprise one or more of:
 a. MSCs;   b. fibroblasts;   c. epithelial cells;   d. stromal cells; and   e. stromal cells and epithelial cells.   
     
     
         9 . The placental product of  claim 8 , wherein the amniotic membrane is substantially free of vascularized tissue or vascularized tissue-derived immunogenic cells. 
     
     
         10 . The placental product of  claim 9 , wherein the amniotic membrane comprises a layer of epithelial cells. 
     
     
         11 . The placental product of  claim 9 , wherein the amniotic membrane comprises one, two, or three layers selected from: a compact layer, a layer of fibroblast, and a spongy layer. 
     
     
         12 . The placental product of  claim 8 , wherein the amniotic membrane is substantially free of immunogenic maternal cells. 
     
     
         13 . The placental product of  claim 8 , wherein the amniotic membrane is substantially free of maternal dendritic cells. 
     
     
         14 . The placental product of  claim 8 , wherein the amniotic membrane is substantially free of maternal leukocytes. 
     
     
         15 . The placental product of any of  claims 8 - 14 , wherein at least: about 40%, or about 50%, or about 60%, or about 70%, or about 74.3%, or about 83.4 or about 90%, or about 92.5% of the stromal cells are viable. 
     
     
         16 . The placental product of  claim 15 , wherein the stromal cells are about 2,000 to about 15,000 in number per cm 2  of the amniotic membrane. 
     
     
         17 . The placental product of any of  claims 15 , wherein the stromal cells are at least: about 2,000, or about 2,400, or about 4,000, or about 6,000, or about 8,000, or about 10,000, or about 10,585, or about 15,000 in number per cm 2 . 
     
     
         18 . The placental product of any of  claims 8 - 14 , wherein about 40% to about 92.5% of the stromal cells are viable. 
     
     
         19 . The placental product of  claim 18 , wherein the stromal cells are about 2,000 to about 15,000 in number per cm 2  of the amniotic membrane. 
     
     
         20 . The placental product of any of  claims 19 , wherein the stromal cells are at least:
 about 2,000, or about 2,400, or about 4,000, or about 6,000, or about 8,000, or about 10,000, or about 10,585, or about 15,000 in number per cm 2 .   
     
     
         21 . The placental product of any of  claims 1 - 14 , wherein the amniotic membrane comprises about 2,000 to about 15,000 stromal cells per cm 2  of the amniotic membrane, optionally, wherein the stromal cells are native to the amniotic membrane. 
     
     
         22 . The placental product of  claim 21 , wherein at least: about 40%, or about 50%, or about 60%, or about 70%, or about 74.3%, or about 83.4 or about 90%, or about 92.5% of the stromal cells are viable. 
     
     
         23 . The placental product of  claim 21 , wherein about 40% to about 92.5% of the stromal cells are viable. 
     
     
         24 . The placental product of any of  claims 1 - 14 , wherein the amniotic membrane comprises at least: about 2,000, or about 2,400, or about 4,000, or about 6,000, or about 8,000, or about 10,000, or about 10,585, or about 15,000 stromal cells per cm 2  of the amniotic membrane, optionally, wherein the stromal cells are native to the amniotic membrane. 
     
     
         25 . The placental product of  claim 24 , wherein at least: about 40%, or about 50%, or about 60%, or about 70%, or about 74.3%, or about 83.4 or about 90%, or about 92.5% of the stromal cells are viable. 
     
     
         26 . The placental product of  claim 24 , wherein about 40% to about 92.5% of the stromal cells are viable. 
     
     
         27 . The placental product of any  claims 1 - 14 , wherein the amniotic membrane comprises about fibroblasts in about 10% to about 30% fibroblasts per total cells. 
     
     
         28 . The placental product of any of  claims 1 - 14 , wherein the amniotic membrane comprises fibroblasts in about 2% to about 30% fibroblasts per total cells. 
     
     
         29 . The placental product of any of  claims 1 - 28 , wherein the placental product or amniotic membrane secretes between about 0 to about 420 pg/mL TNF-α 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 tissue product to a bacterial lipopolysaccharide for about 20 to about 24 hours. 
     
     
         30 . The placental product of  claim 29 , wherein said about between about 0 to about 420 pg/mL TNF-α is: about 0 to about 350 pg/mL TNF-α or 0 to about 280 pg/mL TNF-α. 
     
     
         31 . The placental product of any of  claims 1 - 14 , wherein the amniotic membrane further comprises one or more native factors consisting of the group of IGFBP1, adiponectin, α2-macroglobulin, bFGF, EGF, MMP-9 and TIMP1;
 wherein natives factors are present in an amount:
 i. substantially greater per gram of amniotic membrane than that of a fresh unprocessed amniotic membrane; 
 ii. substantially greater per native amniotic membrane cell than that of a fresh unprocessed amniotic membrane; or 
 iii. substantially greater per μm of amniotic membrane. 
 
 
     
     
         32 . The placental product of any of  claims 1 - 14 , wherein the amniotic membrane has a thickness of about 20 μm to about 50 μm. 
     
     
         33 . The placental product of any of  claims 1 - 14 , wherein the amniotic membrane comprises a basement membrane. 
     
     
         34 . The placental product of any of  claims 1 - 14 , further comprising a chorionic membrane, optionally wherein the chorionic membrane is substantially free of trophoblasts. 
     
     
         35 . A placental product comprising: a cryopreservation medium and an amniotic membrane, wherein:
 a. the amniotic membrane is substantially free of functional CD14+ macrophages; and   b. the cryopreservation medium comprises a cryopreserving amount of a cryopreservative; and   optionally, wherein the amniotic membrane is substantially free of vascularized tissue or vascularized tissue-derived immunogenic cells.   
     
     
         36 . The placental product of  claim 35 , wherein the amniotic membrane comprises viable therapeutic cells, optionally, wherein the viable therapeutic cells are native to the amniotic membrane and of one or more of the types:
 a. MSCs;   b. fibroblasts;   c. epithelial cells;   d. stromal cells and   e. stromal cells and epithelial cells.   
     
     
         37 . The placental product of  claim 36 , wherein the cryopreservative comprises a cell-permeating cryopreservative, a non-cell-permeating cryopreservative, or a combination thereof, optionally wherein the permeating cryopreservative comprises:
 a. DMSO; or   b. DMSO in a majority amount.   
     
     
         38 . The placental product of  claim 37 , wherein the cell-permeating cryopreservative:
 a. does not comprise glycerol in a majority amount; or   b. does not comprises a substantial amount of glycerol.   
     
     
         39 . The placental product of  claim 37 , wherein the cell-permeating cryopreservative comprises a DMSO in amount of: about 2% to about 20%, about 5% to about 20%, about 5% to about 15%, or about 7% to about 13%. 
     
     
         40 . The placental product of  claim 37 , wherein the cryopreservation medium is present in an amount of at least: about 20 ml, about 30 ml, about 35 ml, about 40 ml, about 45 ml, or about 50 ml. 
     
     
         41 . The placental product of  claim 39 , wherein the cryopreservation medium is present in an amount of at least: about 20 ml, about 30 ml, about 35 ml, about 40 ml, about 45 ml, or about 50 ml. 
     
     
         42 . The placental product of  claim 37 , wherein the cryopreservation medium further comprises albumin, optionally wherein the albumin is HSA. 
     
     
         43 . The placental product of  claim 39 , wherein the cryopreservation medium further comprises albumin, optionally wherein the albumin is HSA. 
     
     
         44 . The placental product of any of  claims 36 - 43 , wherein the cryopreservative comprises a non-cell-permeating cryopreservative selected from is a polyvinylpyrrolidone, a starch, a hydroxyethyl starch, a polysacharide, a monosaccharides, a sugar alcohol, an alginate, a trehalose, a raffinose, a dextran, or a combination thereof. 
     
     
         45 . The placental product of any of  claims 36 - 43 , wherein at least: about 40%, or about 50%, or about 60%, or about 70%, or about 74.3%, or about 83.4 or about 90%, or about 92.5% of the stromal cells are viable after a freeze-thaw cycle. 
     
     
         46 . The placental product of  claim 45 , wherein the stromal cells are about 2,000 to about 15,000 in number per cm 2  of the amniotic membrane. 
     
     
         47 . The placental product of  claim 45 , wherein the stromal cells are at least: about 2,000, or about 2,400, or about 4,000, or about 6,000, or about 8,000, or about 10,000, or about 10,585, or about 15,000 in number per cm 2 . 
     
     
         48 . The placental product of any of  claims 36 - 43 , wherein about 40% to about 92.5% of the stromal cells are viable after a freeze-thaw cycle. 
     
     
         49 . The placental product of  claim 48 , wherein the stromal cells are about 2,000 to about 15,000 in number per cm 2  of the amniotic membrane. 
     
     
         50 . The placental product of any of  claims 48 , wherein the stromal cells are at least:
 about 2,000, or about 2,400, or about 4,000, or about 6,000, or about 8,000, or about 10,000, or about 10,585, or about 15,000 in number per cm 2 .   
     
     
         51 . The placental product of any of  claims 35 - 43 , wherein the amniotic membrane comprises about 5,000 to about 50,000 stromal cells per cm 2  of the amniotic membrane, optionally, wherein the stromal cells are native to the amniotic membrane. 
     
     
         52 . The placental product of  claim 51 , wherein at least: about 40%, or about 50%, or about 60%, or about 70%, or about 74.3%, or about 83.4 or about 90%, or about 92.5% of the stromal cells are viable after a freeze-thaw cycle. 
     
     
         53 . The placental product of  claim 51 , wherein about 40% to about 92.5% of the stromal cells are viable. 
     
     
         54 . The placental product of any of  claims 35 - 43 , wherein the amniotic membrane comprises at least: about 2,000, or about 2,400, or about 4,000, or about 6,000, or about 8,000, or about 10,000, or about 10,585, or about 15,000 stromal cells per cm 2  of the amniotic membrane, optionally, wherein the stromal cells are native to the amniotic membrane. 
     
     
         55 . The placental product of  claim 54 , wherein at least: about 40%, or about 50%, or about 60%, or about 70%, or about 74.3%, or about 83.4 or about 90%, or about 92.5% of the stromal cells are viable after a freeze-thaw cycle. 
     
     
         56 . The placental product of  claim 54 , wherein about 40% to about 92.5% of the stromal cells are viable after a freeze-thaw cycle. 
     
     
         57 . The placental product of any  claims 35 - 43 , wherein the amniotic membrane comprises fibroblasts in about 2% to about 30% fibroblasts per total cells. 
     
     
         58 . The placental product of any of  claims 35 - 43 , wherein the amniotic membrane comprises CD14+ macrophage at less than about 10% or less than about 2% or less than about 0.5% of the total cells. 
     
     
         59 . The placental product of any of  claims 35 - 43 , wherein the amniotic membrane is substantially free of:
 a. maternal dendritic cells, maternal leukocytes, or a combination thereof, or   b. immunogenic maternal cells,   
     
     
         60 . The placental product of any of  claims 35 - 43 , wherein the product comprises about 2,000 to about 15,000 stromal cells per cm 2  and wherein the viability of said stromal cells after a freeze-thaw cycle is about 70% to about 95% relative to fresh placental or amniotic membrane. 
     
     
         61 . The placental product of any of  claims 35 - 43 , wherein after a freeze-thaw cycle, the placental product or amniotic membrane secretes between about 0 to about 420 pg/mL TNF-α 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 tissue product to a bacterial lipopolysaccharide for about 20 to about 24 hours. 
     
     
         62 . The placental product of any of  claims 35 - 43 , wherein after a refrigeration-freeze-thaw cycle, the placental product or amniotic membrane secretes between about 0 to about 420 pg/mL TNF-α 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 tissue product to a bacterial lipopolysaccharide for about 20 to about 24 hours. 
     
     
         63 . The placental product of  claim 61 , wherein said about between about 0 to about 420 pg/mL TNF-α is: about 0 to about 350 pg/mL TNF-α or 0 to about 280 pg/mL TNF-α. 
     
     
         64 . The placental product of any of  claims 35 - 43 , wherein the amniotic 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:
 i. substantially greater per gram of amniotic membrane than that of a fresh unprocessed amniotic membrane; 
 ii. substantially greater per native amniotic membrane cell than that of a fresh unprocessed amniotic membrane; or 
 iii. substantially greater per μm of amniotic membrane. 
 
     
     
         65 . The placental product of any of  claims 35 - 43 , wherein the amniotic membrane has a thickness of about 20 μm to about 50 μm. 
     
     
         66 . The placental product of any of  claims 35 - 43 , further comprising a chorionic membrane, optionally wherein the chorionic membrane is substantially free of trophoblasts. 
     
     
         67 . The placental product of  claim 66 , wherein the amniotic membrane and the chorion membrane are associated to each another in the native architecture. 
     
     
         68 . The placental product of  claim 66 , wherein the amniotic membrane and the chorionic membrane are not attached to one another in the native architecture. 
     
     
         69 . The placental product of any of  claims 35 - 43 , wherein the amniotic membrane comprises a layer of epithelial cells. 
     
     
         70 . The placental product of any of  claims 35 - 43 , wherein the amniotic membrane comprises one, two, or three layers selected from: a compact layer, a layer of fibroblast, and a spongy layer. 
     
     
         71 . The placental product of any of  claims 35 - 43 , wherein the amniotic membrane has a thickness of about 20 μm to about 50 μm. 
     
     
         72 . The placental product of any of  claims 35 - 43 , wherein the amniotic membrane comprises a layer of epithelial cells. 
     
     
         73 . The placental product of any of  claims 35 - 43 , wherein the amniotic membrane comprises a basement membrane. 
     
     
         74 . A placental product comprising: a cryopreservation medium and an amniotic membrane, wherein:
 a. the amniotic membrane:
 i. comprises a compact layer, layer of fibroblast, and a spongy layer, wherein each of said layers comprises viable therapeutic cells; and 
 ii. is substantially free of vascularaized tissue and functional CD14+ macrophages; and 
   b. the cryopreservation medium comprises a cryopreserving amount of a cryopreservative.   
     
     
         75 . The placental product of  claim 74 , wherein the amniotic membrane comprises about 2,000 to about 15,000 stromal cells per cm 2  of the amniotic membrane. 
     
     
         76 . The placental product of  claim 75 , wherein at least: about 40%, or about 50%, or about 60%, or about 70%, or about 74.3%, or about 83.4 or about 90%, or about 92.5% of the stromal cells are viable. 
     
     
         77 . The placental product of any  claim 75 , wherein about 40% to about 92.5% of the stromal cells are viable. 
     
     
         78 . The placental product of  claim 74 , wherein the amniotic membrane comprises at least: about 2,000, or about 2,400, or about 4,000, or about 6,000, or about 8,000, or about 10,000, or about 10,585, or about 15,000 stromal cells per cm 2  of the amniotic membrane. 
     
     
         79 . The placental product of  claim 75 , wherein at least: about 40%, or about 50%, or about 60%, or about 70%, or about 74.3%, or about 83.4 or about 90%, or about 92.5% of the stromal cells are viable after a freeze-thaw cycle. 
     
     
         80 . The placental product of  claim 75 , wherein about 40% to about 92.5% of the stromal cells are viable after a freeze-thaw cycle. 
     
     
         81 . The placental product of any of  claims 74 - 80 , wherein the amniotic membrane further comprises a basement membrane. 
     
     
         82 . The placental product of any of  claims 74 - 81 , wherein the amniotic membrane has a thickness of about 20 μm to about 50 μm. 
     
     
         83 . The placental product of any of  claims 74 - 82 , wherein the cryopreservative comprises DMSO. 
     
     
         84 . The placental product of any of  claims 1 - 14 ,  35 - 43 , or  74 - 80 , wherein the amniotic membrane comprises MSCs in an amount of: at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, about 1% to about 10%, or about 3% to about 10%, relative to the total number of cells in the amniotic membrane. 
     
     
         85 . The placental product of  claim 84 , wherein at least about 40%, about 50%, about 60%, or about 70% of the MSCs are viable. 
     
     
         86 . The placental product of any of  claims 1 - 14 ,  35 - 43 , or  74 - 80 , wherein the amniotic membrane comprises fibroblasts in an amount of: about 1%, about 20%, about 5% to about 15%, at least about 1%, at least about 2%, at least about 3%, or at least about 4 relative to the total number of cells in the amniotic membrane. 
     
     
         87 . The placental product of  claim 86 , wherein at least: about 40%, about 50%, about 60%, or about 70% of the fibroblasts are viable. 
     
     
         88 . The placental product of any of  claims 1 - 14 ,  35 - 43 , or  74 - 80 , wherein the amniotic membrane comprises stromal cells in an amount of: about 5% to about 40%, about 5% to about 30%, about 10% to about 30%, about 15% to about 25%, at least about 5%, at least about 10%, or at least about 15%, relative to the total number of cells in the amniotic membrane. 
     
     
         89 . The placental product of  claim 88 , wherein at least: about 40%, about 50%, about 60%, or about 70% of the stromal cells are viable after a freeze-thaw cycle 
     
     
         90 . The placental product of any of  claims 1 - 14 ,  35 - 43 , or  74 - 80 , wherein the amniotic membrane comprises epithelial cells in an amount of: about 60% to about 90%, about 70% to about 90%, about 40% to about 90%, about 50% to about 90%, at least about 40%, at least about 50%, least about 60%, or at least about 70%, relative to the total number of cells in the amniotic membrane. 
     
     
         91 . The placental product of  claim 90 , wherein at least: about 40%, about 50%, about 60%, or about 70% of the epithelial cells are viable after a freeze-thaw cycle. 
     
     
         92 . The placental product of any of  claims 1 - 14 ,  35 - 43 , or  74 - 80 , wherein the amniotic membrane comprises MSCs and functional macrophages in a ratio of greater than about any of: 5:1, 7:1, or 10:1. 
     
     
         93 . The placental product of any of  claims 1 - 14 ,  35 - 43 , or  74 - 80 , wherein the amniotic membrane comprises fibroblasts and functional macrophages in a ratio of greater than about any of: 10:1, 15:1, 20:1, or 25:1. 
     
     
         94 . The placental product of any of  claims 1 - 14 ,  35 - 43 , or  74 - 80 , wherein the amniotic membrane comprises fibroblasts and MSCs in a ratio of: about 4:1 to about 1:1 or about 3:1 to about 3:2, or about 2:1. 
     
     
         95 . The placental product of any of  claims 1 - 14 ,  35 - 43 , or  74 - 80 , wherein the amniotic membrane comprises MSCs in an amount of: at least about 1,000 cells/cm 2 ′ at least about 2,000 cells/cm 2 , about 1,000 to about 5,000 cells/cm 2 , or about 2,000 to about 5,000 cells/cm 2 . 
     
     
         96 . The placental product of  claim 95 , wherein at least: about 40%, about 50%, about 60%, or about 70% of the MSCs are viable after a freeze-thaw cycle 
     
     
         97 . The placental product of any of  claims 1 - 14 ,  35 - 43 , or  74 - 80 , wherein the amniotic membrane comprises fibroblasts in an amount of: at least about 2,000 cells/cm 2 , at least about 4,000 cells/cm 2 , about 2,000 to about 9,000 cells/cm 2  , or about 2,000 to about 9,000 cells/cm 2 . 
     
     
         98 . The placental product of  claim 97 , wherein at least: about 40%, about 50%, about 60%, or about 70% of the fibroblasts are viable after a freeze-thaw cycle 
     
     
         99 . The placental product of any of  claims 1 - 14 ,  35 - 43 , or  74 - 80 , wherein the amniotic membrane comprises stromal cells in an amount of: at least about 4,000, at least about 8,000 cells/cm 2 , about 4,000 to about 18,000 cells/cm 2 , or about 4,000 to about 18,000 cells/cm 2 . 
     
     
         100 . The placental product of  claim 99 , wherein at least: about 40%, about 50%, about 60%, or about 70% of the stromal cells are viable after a freeze-thaw cycle. 
     
     
         101 . The placental product of any of  claims 1 - 14 ,  35 - 43 , or  74 - 80 , wherein the amniotic membrane comprises epithelial cells in an amount of: at least about 10,000 cells/cm 2 , at least about 20,000 cells/cm 2 , at least about 32,000 cells/cm 2 , about 10,000 to about 72,000 cells/cm 2  , about 20,000 to about 72,000 cells/cm 2 , or about 32,000 to about 72,000 cells/cm 2    
     
     
         102 . The placental product of  claim 101 , wherein at least: about 40%, about 50%, about 60%, or about 70% of the epithelial cells are viable after a freeze-thaw cycle. 
     
     
         103 . The placental product of any of  claims 1 - 14 ,  35 - 43 , or  74 - 80 , wherein the amniotic membrane comprises functional macrophages in an amount of: less than about 3,000 cells/cm 2 , less than about 1,000 cells/cm 2 , or less than about 500 cells/cm 2 . 
     
     
         104 . A method of treating a wound of a subject comprising administering the placental product of any of the preceding Claims to the wound. 
     
     
         105 . The method of  claim 94 , wherein the wound is an ulcer. 
     
     
         106 . The method of  claim 95 , wherein the ulcer is a foot ulcer, optionally a diabetic foot ulcer. 
     
     
         107 . The method of  claim 95 , wherein the ulcer is a diabetic foot ulcer. 
     
     
         108 . The method of any of  claims 93 - 97 , wherein the subject has diabetes. 
     
     
         109 . A method of ligament or tendon surgery comprising applying the placental product of any of  claims 1 - 103  to a ligament or tendon. 
     
     
         110 . The method of  claim 109 , wherein the ligament or tendon surgery is a grafting surgery, a reconstruction surgery, or a repair of a torn ligament or tendon. 
     
     
         111 . The method of  claim 110 , wherein the grafting surgery comprises the attachment of a graft to a biological substrate, optionally, wherein the biological substrate is bone. 
     
     
         112 . The method of  claim 111 , wherein the graft is an allograft, a xenograft, or an autologous graft. 
     
     
         113 . The method of  claim 110 , wherein the ligament or tendon surgery is selected from: knee surgery, shoulder surgery, leg surgery, and arm surgery, elbow surgery, finger surgery, hand surgery, wrist surgery, toe surgery, foot surgery, ankle surgery. 
     
     
         114 . The method of  claim 110 , wherein the tendon is hamstring tendon, a bicep tendon, an Achilles Tendon, an extensor tendon, or a rotator cuff tendon. 
     
     
         115 . The method of  claim 110 , wherein the step of applying the placental product comprises: suturing the placental product to the tendon or ligament, enveloping the tendon or ligament with the placental product, or a combination thereof. 
     
     
         116 . A method of tissue grafting comprising applying the placental product of any of  claims 1 - 103  to a tissue graft, and attaching the tissue graft to a biological substrate. 
     
     
         117 . The method of  claim 116 , wherein the placental product is applied to a portion of the tissue graft which is attached to the biological substrate. 
     
     
         118 . The method of  claim 117 , wherein the step of applying the placental product comprises: suturing the placental product to the graft, enveloping graft with the placental product, or a combination thereof. 
     
     
         119 . The method of  claim 118 , wherein the tissue graft comprises a tissue selected from: skin, cartilage, ligament, tendon, periosteum, perichondrium, synovium, fascia, and mesenter. 
     
     
         120 . A method of inhibiting fibrosis or adhesion at a wound site comprising applying the placental product of any of  claims 1 - 103  to the wound site. 
     
     
         121 . The method of  claim 120 , wherein the wound site is surgery induced or associated with surgery. 
     
     
         122 . The method of  claim 121  wherein the surgery is a surgery selected from: spine, laminectomy, knee, shoulder, or child birth, eye, cardiovascular, angiogenesis stimulation, burn, and brain/neurological procedures. 
     
     
         123 . The method of  claim 120 , wherein the wound site is a trauma related wound. 
     
     
         124 . A method of treating a tissue injury of a subject comprising administering the placental product of any of  claims 1 - 103  to the tissue injury. 
     
     
         125 . The method of  claim 124 , wherein the tissue injury is an ulcer. 
     
     
         126 . The method of  claim 125 , wherein the ulcer is a foot ulcer, optionally a diabetic foot ulcer. 
     
     
         127 . The method of  claim 126 , wherein the ulcer is a diabetic foot ulcer. 
     
     
         128 . The method of any of  claims 125  wherein the subject has diabetes. 
     
     
         129 . The method of any of  claims 125  wherein the wound is associated with ischemia. 
     
     
         130 . A method of promoting migration or proliferation of at least one cell selected from the group consisting of fibroblasts, epithelial cells, and endothelial cells] in a subject comprising administering the placental product of any of  claims 1 - 103  a subject in need thereof. 
     
     
         131 . A method of reorganizing extracellular matrix of a subject comprising administering the placental product of any of  claims 1 - 103  to the subject. 
     
     
         132 . A method of treating a wound comprising providing at least one factor that is a chemotactic factor or a growth factor to the wound by administering the placental product of any of  claims 1 - 103  to the subject. 
     
     
         133 . A method of providing stem cells to tissue of a subject comprising administering the placental product of any of  claims 1 - 103  to the subject.