Composite for Thermo-Sensitive Cell-Tissue Transplanted Scaffold and Use thereof

Abstract

A composite comprising a stem cell; a biodegradable layer, which can provide an environment for the stem cell to grow and to differentiate, and; a N-isopropylacrylamide (NIPAAm), which can polymerize with the biodegradable layer and possess the temperature-responsive character for easy stripping. This invention also provides a method of treating a subject with a skin defect by covering the composite of the present invention on the skin defect of the subject in need of such treatment. Furthermore, using this composite with different growth factors, stem cells can be induced to differentiate into skin-related, neuronal cells, neuron, and insulin-positive cells in biodegradable scaffolds as well as transplanted graft. Finally, this invention also provides a quick and convenient method of monitoring cell growth or tissue engineering in an animal.

1 . A composite comprising:
 (a) a stem cell which differentiates into mature normal cell, osteocyte, chondrocyte, adipocyte, epithelium cell, epidermis-related cell, keratocyte, neuron, neuronal cell, insulin-positive cell, glucagon-positive cell, or tissues thereof;   (b) a biodegradable layer for the stem cell to grow and differentiation, wherein the layer contains material selected from the group consisting of gelatin, fibronectin, collagen, laminin, bFGF, EGF, insulin, progesterone, glucose, SDF and thymosin beta-4; and   (c) a N-isopropylacrylamide (NIPAAm), which polymerizes with the biodegradable layer to provide the feature of thermo-sensitive response for easy stripping.   
     
     
         2 . The composition of  claim 1 , wherein the biodegradable layer is selected from the group consisting of gelatin, fibronectin, collagen, laminin, bFGF and EGF. 
     
     
         3 . The composition of  claim 1 , wherein the stem cell is selected from the group consisting of (a) adult stem cell derived from bone marrow, umbilical tissues, or placenta; (b) neural stem cell; and (c) embryomic stem cell. 
     
     
         4 . The composition of  claim 1 , wherein the N-isopropylacrylamide (NIPAAm) further polymerizes with a cover. 
     
     
         5 . The composition of  claim 4 , wherein the cover is PP non-woven. 
     
     
         6 . A method for preparing a composite of  claim 1  comprising:
 (a) irradiating a NIPAAm solution with a UV light;   (b) crosslinking the NIPAAm-grafted a cover in gelatin solution by the glutaraldehyde crosslinking agent;   (c) freezing and drying the NIPAAm-grafted cover;   (d) immersing the dried NIPAAm-grafted cover into the glutaraldehyde solution to produce a crosslinked gelatin hydrogel;   (e) treating the crosslinked gelatin hydrogel with a glycine aqueous solution to block non-reacted aldehyde groups, and;   (f) cultivating a stem cell on the crosslinked gelatin hydrogel.   
     
     
         7 . The method of  claim 6 , wherein the biodegradable layer is selected from the group consisting of gelatin, fibronectin, collagen, laminin, bFGF and EGF. 
     
     
         8 . The method of  claim 6 , wherein the stem cell is selected from the group consisting of (a) adult stem cell derived from bone marrow, umbilical tissues, or placenta; (b) neural stem cell; and (c) embryomic stem cell. 
     
     
         9 . The method of  claim 6 , wherein the UV light is 50-2000 W UV light. 
     
     
         10 . The method of  claim 6 , wherein NIPAAm-grafted PP non-woven in step (c) is dried out. 
     
     
         11 . The method of  claim 6 , which further washes with the double-distilled water after step (e). 
     
     
         12 . The method of  claim 6 , wherein the cover is a PP non-woven. 
     
     
         13 . A method of treating a subject with a skin defect comprises covering the composite of  claim 1  on the skin defect of the subject in need of such treatment. 
     
     
         14 . The method of  claim 13  wherein the skin defect is burn, trauma or wound. 
     
     
         15 . The method of  claim 14  wherein the wound is resulted from a surgery or a burn. 
     
     
         16 . The method of  claim 13  wherein the N-isopropylacrylamide (NIPAAm) further polymerizes with a cover. 
     
     
         17 . The method of  claim 16 , wherein the cover is easily peeled by treating under room temperature on the cover. 
     
     
         18 . A method of monitoring cell growth or tissue engineer in an animal comprising applying the composite to the animal in need of such monitoring, wherein the composite comprises a stem cell labeled by a marker; a biodegradable layer, which can provide an environment for the stem cell to grow, differentiate, and; a N-isopropylacrylamide (NIPAAm), which can polymerize with the biodegradable layer. 
     
     
         19 . The method of  claim 18 , wherein the monitoring is directed to development induction of skin-related, pancreas islet-related cells, neural cell, or neurons. 
     
     
         20 . The method of  claim 19  wherein the pancreas islet-related cells is like insulin- and glucagon-positive cells. 
     
     
         21 . The method of  claim 18 , wherein the marker is green, red or blue fluorescent protein gene.