Membrane and method for culture and differentiation of cells

Abstract

Provided is a membrane for cell culture and differentiation. The membrane has a base portion and an array of protrusions consisting of a plurality of protrusions. The protrusions are substantially evenly distributed on the base portion. The plurality of protrusions has dimensions on the order of micrometers. In particular, the membrane consists of particles of different particle sizes of two or more kinds. One kind of particles have an average particle size of 1 μm to 50 μm. Two or more kinds of particles of different particle sizes include nanoscale particles, 10-900 nm. One kind of particles are selected from the group consisting of inorganic compound microspheres. The other kind of particles of the two or more kinds of particles of different particle sizes are selected from the group consisting of organic polymer nanospheres. Also provided is a method for maintaining, culturing and/or differentiating cells using such membrane.

What is claimed is: 
     
         1 . A membrane for culture and differentiation of cells, comprising:
 (A) a base portion; and   (B) a protrusion array composed of a plurality of protrusions, the plurality of protrusions being substantially uniformly distributed on the base portion, and the plurality of protrusions having a size on the order of micrometers.   
     
     
         2 . The membrane according to  claim 1 , wherein a distance between adjacent protrusions is on the order of micrometers. 
     
     
         3 . The membrane according to  claim 1 , wherein the membrane is a colloidal crystal membrane. 
     
     
         4 . The membrane according to  claim 1 , wherein the membrane is composed of particles of different particle sizes of two or more kinds, and the two or more kinds of particles of different particle sizes comprise at least:
 particles of a first kind, serving as the protrusions and having an average particle size of 1 μm to 50 μm, and   particles of a second kind, serving as the base portion and having an average particle size less than or equal to ½ of the average particle size of the first kind of particles.   
     
     
         5 . The membrane according to  claim 1 , wherein the membrane is composed of particles of different particle sizes of two kinds, wherein in the particles of different particle sizes of the two kinds, particles of a first kind have an average particle size of 1 μm to 50 μm, and particles of a second kind have an average particle size of 10-900 nm. 
     
     
         6 . The membrane according to  claim 5 , wherein the particles of the first kind are made of an inorganic compound. 
     
     
         7 . The membrane according to  claim 5 , wherein the particles of the second kind are made of an organic polymer. 
     
     
         8 . The membrane according to  claim 5 , wherein the particles of the first kind are made of one or more materials selected from the group consisting of silica, titania, zinc oxide, chemically modified silica, chemically modified titanium dioxide, chemically modified zinc oxide, and any combination thereof. 
     
     
         9 . The membrane according to  claim 5 , wherein the particles of the second kind are made of one or more materials selected from the group consisting of polystyrene, acrylic polymers, chitosan, poly(lactic-co-glycolic acid), polylactic acid, polycaprolactone, gelatin and any combination thereof. 
     
     
         10 . The membrane according to  claim 9 , wherein the acrylic polymers are selected from the group consisting of poly(meth)acrylic acids, poly(meth)acrylates and any combination thereof. 
     
     
         11 . The membrane according to  claim 10 , wherein the poly(meth)acrylates are selected from the group consisting of poly(meth)acrylic acid C 1 -C 20  alkyl esters. 
     
     
         12 . The membrane according to  claim 10 , wherein the poly(meth)acrylates are selected from the group consisting of poly(methyl acrylate), poly(methyl methacrylate), poly(ethyl acrylate), poly(ethyl methacrylate), poly(propyl acrylate), poly(propyl methacrylate), poly(butyl acrylate), poly(butyl methacrylate), poly(pentyl acrylate), poly(pentyl methacrylate), poly(hexyl acrylate), poly(hexyl methacrylate) and any combination thereof. 
     
     
         13 . The membrane according to  claim 5 , wherein the particles of the first kind are made of silica and the particles of the second kind are made of poly(methyl methacrylate). 
     
     
         14 . The membrane according to  claim 5 , wherein the particles of the first kind are in form of a single layer of particles, and a ratio of the particles of the first kind and the particles of the second kind is set such that the particles of the first kind are distributed in the particles of the second kind in a partially embedded manner to form a colloidal crystal. 
     
     
         15 . A method for culture and/or differentiation of cells, comprising culturing and/or differentiating the cells on the membrane according to  claim 1 . 
     
     
         16 . The method according to  claim 15 , wherein the cells are selected from the group consisting of induced pluripotent stem cells, embryonic stem cells and adult stem cells. 
     
     
         17 . The method according to  claim 15 , wherein the cells are selected from the group consisting of bone marrow mesenchymal stem cells, hematopoietic stem cells, neural stem cells, peripheral blood stem cells, adipose stem cells, placental stem cells, placental sub-totipotent stem cells, and amniotic stem cells. 
     
     
         18 . The method according to  claim 15 , wherein the cells are human induced pluripotent stem cells, and by the method, the human induced pluripotent stem cells are differentiated in a directed differentiation manner into human induced pluripotent stem cell-derived cardiomyocytes. 
     
     
         19 . The method according to  claim 15 , further comprising separating the cells from the membrane by liquid flushing or suction after the differentiation. 
     
     
         20 . A method for maintaining growth and stemness of cells, comprising
 culturing the cells on the membrane according to  claim 1 ,   wherein the cells are selected from the group consisting of induced pluripotent stem cells, embryonic stem cells and adult stem cells.