Production of extracellular matrix, conditioned media and uses thereof

Abstract

Provided is a matrix for promoting survival and differentiation of cells transplanted thereon, comprising a base matrix and a cell-made matrix thereon. Methods and means for making and using same are also provided. Also provided are conditioned media, related compositions, related methods, and related packaging products.

1 . A method of increasing survival and/or differentiation of target cells on a base matrix, the method comprising:
 creating a cell-made extracellular matrix on said base matrix to produce a modified base matrix and   administering said target cells to said modified base matrix.   
     
     
         2 . The method of  claim 1 , wherein the step of creating the cell-made extracellular matrix on said base matrix to produce the modified base matrix is performed in vitro. 
     
     
         3 . The method of  claim 1 , wherein the step of administering said target cells to said modified base matrix is performed in vivo. 
     
     
         4 . The method of  claim 1 , wherein the step of creating the cell-made extracellular matrix on said base matrix to produce the modified base matrix is performed in vivo. 
     
     
         5 . A modified base matrix for survival and/or differentiation of target cells thereon, the modified base matrix comprising a cell-made extracellular matrix thereon. 
     
     
         6 . The modified base matrix of  claim 5 , wherein the modified base matrix is produced by culturing cells capable of forming said cell-made extracellular matrix on the base matrix. 
     
     
         7 . The modified base matrix of  claim 5 , wherein said modified base matrix is produced by treating the base matrix with at least an active fraction of a conditioned media from culturing cells capable of forming said cell-made extracellular matrix on the base matrix. 
     
     
         8 . The modified matrix of  claim 5 , wherein said base matrix is Bruch's membrane. 
     
     
         9 . The modified base matrix of  claim 8 , wherein the Bruch's membrane is undergoing macular degeneration. 
     
     
         10 . The modified matrix of  claim 5 , wherein said base matrix is a synthetic polymer-based matrix. 
     
     
         11 . The modified base matrix of  claim 10 , wherein the synthetic polymer is selected from polylactic acid (PLA), polyglycolic acid (PGA), poly(lactide-co-glycolide) (PLGA), poly(methyl methacrylate) (PMMA), polyorthoesters, and any combinations thereof. 
     
     
         12 . The modified base matrix of  claim 11 , wherein the synthetic polymer is polycaprolactone (PCL). 
     
     
         13 . The modified base matrix of  claim 5 , wherein said target cells are selected from RPE, umbilical cells, placental cells, adult stem cells, embryonic stem cells, fetal RPE, adult iris pigment epithelial (IPE) cells, bone marrow-derived stem cells, Schwann cells, neural progenitor cells, and any combination thereof. 
     
     
         14 . The modified base matrix of  claim 5 , wherein said target cells are autologous. 
     
     
         15 . The modified base matrix of  claim 5 , wherein said target cells are fetal RPEs. 
     
     
         16 . The modified base matrix of  claim 5 , wherein the target cells are adult or embryonic stem cells or are differentiated from adult or embryonic stem cells. 
     
     
         17 . A conditioned media from culturing cells capable of forming a cell-made extracellular matrix on the base matrix of  claim 5 . 
     
     
         18 . The conditioned media of  claim 17 , which is serum-free. 
     
     
         19 . The conditioned media of  claim 17 , wherein said cells capable of forming said cell-made extracellular matrix on the base matrix are selected from corneal endothelial cells, RPE cells, IPE cells, embryonic stem cells, bone marrow-derived stem cells, placental cells, and/or umbilical cells. 
     
     
         20 . The conditioned media of  claim 19 , wherein said cells are corneal endothelial cells. 
     
     
         21 . The conditioned media of  claim 19 , wherein the conditioned media or at least a fraction thereof is produced by culturing cells capable of forming said cell-made extracellular matrix. 
     
     
         22 . An active fraction of a conditioned media of  claim 17 , characterized by a depletion of biologically active components having MW of less than about 100 kD. 
     
     
         23 . The active fraction of  claim 22 , wherein the active fraction of the conditioned media from culturing cells capable of forming said cell-made extracellular matrix on the base matrix is formed by a depletion of biologically active components having MW of less than about 100 kD. 
     
     
         24 . A kit for promoting the survival and/or differentiation of target cells on a base matrix, comprising:
 a) a set of instructions and at least one of:   b) an efficient amount of cells capable of forming a cell-made extracellular matrix; and   c) at least an active fraction of a conditioned media or an extracellular matrix from the cells capable of forming a cell-made extracellular matrix.   
     
     
         25 . The kit of  claim 24 , wherein the cells capable of forming the cell base and at least the active fraction of the conditioned media or the extracellular matrix from the cells capable of forming the cell-made extracellular matrix are according  claim 5 . 
     
     
         26 . A kit for promoting survival and differentiation of target cells comprising a set of instructions and a modified base matrix according to  claim 5 . 
     
     
         27 . The kit of  claim 24  further comprising an effective amount of target cells selected from RPE, umbilical cells, placental cells, adult stem cells, cells differentiated from adult stem cells, ES cells, cells differentiated from ES cells, bone marrow-derived stem cells, fetal RPE, adult iris pigment epithelial (IPE) cells, Schwann cells, and any combination thereof, derived from autologous or allogenic source. 
     
     
         28 . A method for making a conditioned medium, comprising
 obtaining a plurality of cells capable of forming a cell-made extracellular matrix;   culturing the cells in a first medium for a period of time to form a second medium; and   collecting the second medium thereby making the conditioned medium.   
     
     
         29 . The method of  claim 28 , wherein the first medium is free of serum. 
     
     
         30 . The method of  claim 28 , wherein the cells are selected from the group consisting of corneal endothelial cells, RPE cells, IPE cells, embryonic stem cells, bone marrow-derived stem cells, placental cells, and umbilical cells. 
     
     
         31 . The method of  claim 30  wherein the cells are corneal endothelial cells. 
     
     
         32 . The method of  claim 28 , wherein the period of time is 1-5 days. 
     
     
         33 . The method of  claim 32 , wherein the period of time is 2-4 days. 
     
     
         34 . The method of  claim 33 , wherein the period of time is 3 days. 
     
     
         35 . A composition comprising a fraction of a conditioned medium of  claim 17 . 
     
     
         36 . The composition of  claim 35 , wherein the fraction comprises molecules having MW of less than 3 kD. 
     
     
         37 . The composition of  claim 35 , wherein the fraction comprises molecules having MW of less than 50 kD. 
     
     
         38 . The composition of  claim 37 , wherein the fraction comprises molecules having MW of 10-50 kD. 
     
     
         39 . The composition of  claim 38 , wherein the fraction comprises molecules having MW of 30-50 kD. 
     
     
         40 . The composition of  claim 35 , comprising a first fraction and a second fraction of the medium, wherein (i) the first fraction comprises molecules having MW of less than 3 kD, and (ii) the second fraction comprises molecules having MW of 10-50 kD. 
     
     
         41 . The composition of  claim 35 , wherein the fraction comprises one or more proteins selected from those in Table 6. 
     
     
         42 . The composition of  claim 35 , wherein the composition is a pharmaceutical composition and comprises a pharmaceutically acceptable carrier. 
     
     
         43 . The composition of  claim 35 , wherein the composition is a cell culture medium for use in storing, preserving, or inducing differentiation of cells or tissues. 
     
     
         44 . A method for treating age-related macular degeneration (AMD), comprising
 identifying a subject in need thereof; and   administering to the subject an effective amount of the composition of  claim 35 .   
     
     
         45 . A packaging product comprising
 a composition of  claim 44 ;   a cell or a piece of tissue, and   a container holding the composition.   
     
     
         46 . The packaging product of  claim 45 , wherein the cell is selected from the group consisting of retinal pigment epithelial (RPE) cells, stem cells, and corneal cells. 
     
     
         47 . The packaging product of  claim 45 , wherein the tissue is selected from the group consisting of RPE derived from precursor cells, RPE derived from human embryonic stem cells, RPE derived from iPSC stem cells, whole retinae, whole cornea, tissues, and neural tissues and organs. 
     
     
         48 . The packaging product of  claim 45 , wherein the cell is in suspension or in a support matrix designed for cell delivery. 
     
     
         49 . The packaging product of  claim 45 , where in the composition has a temperature within the range of 0-40° C. 
     
     
         50 . The packaging product of  claim 49 , wherein the composition has a temperature within the range of 4-30° C. 
     
     
         51 . The packaging product of  claim 50 , wherein the composition has a temperature within the range of 15-25° C. 
     
     
         52 . The packaging product of  claim 44 , wherein the container is sealed.