Membrane and method for culture and differentiation of cells
Provided is a membrane for cell culture and differentiation. The membrane comprises 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 types. One type of particles has an average particle size of 1 μm to 50 μm. Two or more types of particles of different particle sizes include nanoscale particles, 10-900 nm. One type of particle is selected from the group consisting of inorganic compound microspheres. The other type of particles of the two or more types of particles of different particle sizes is selected from the group consisting of organic polymer nanospheres. Also provided is a method for maintaining, culturing and/or differentiating cells using such membrane.
1 . A method for differentiation of human induced pluripotent stem cells (hiPSCs) into hiPSC-derived cardiomyocytes (hiPSC-CMs), comprising differentiating the hiPSCs on a membrane pre-coated with a solubilized basement membrane preparation comprising:
a base portion; and
a protrusion array composed of a plurality of protrusions, the plurality of protrusions being substantially uniformly distributed on the base portion, and, each protrusion having a micrometer-scale dimension.
2 . The method according to claim 1 , further comprises detaching and collecting the cells from the membrane by liquid flushing or suction without using an enzyme.
3 . The method according to claim 1 , wherein a distance between adjacent protrusions is on the order of micrometers.
4 . The method according to claim 1 , wherein the membrane is a colloidal crystal membrane.
5 . The method according to claim 1 , wherein the membrane is composed of particles of different particle sizes of two or more types, and the two or more types of particles of different particle sizes comprise at least:
particles of a first type, serving as the protrusions and having an average particle size of 1 μm to 50 μm, and
particles of a second type, serving as the base portion and having an average particle size less than or equal to 1/10 of the average particle size of the first type of particles.
6 . The method according to claim 1 , wherein the membrane is composed of particles of different particle sizes of two types, wherein in the particles of different particle sizes of the two types, particles of a first type have an average particle size of 1 μm to 50 μm, and particles of a second type have an average particle size of 10-900 nm.
7 . The method according to claim 6 , wherein the particles of the first type are made of an inorganic compound.
8 . The method according to claim 6 , wherein the particles of the second type are made of an organic polymer.
9 . The method according to claim 6 , wherein the particles of the first type 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.
10 . The method according to claim 6 , wherein the particles of the second type 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.
11 . The method according to claim 10 , wherein the acrylic polymers are selected from the group consisting of poly(meth)acrylic acids, poly(meth)acrylates and any combination thereof.
12 . The method according to claim 11 , wherein the poly(meth)acrylates are selected from the group consisting of poly(meth)acrylic acid C 1 -C 20 alkyl esters.
13 . The method according to claim 11 , wherein the poly(meth)acrylates comprise at least one 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.
14 . The method according to claim 6 , wherein the particles of the first type are made of silica and the particles of the second type are made of poly(methyl methacrylate).
15 . The method according to claim 6 , wherein the particles of the first type are in the form of a single layer of particles, and a ratio of the particles of the first type and the particles of the second type is set such that the particles of the first type are distributed in the particles of the second type in a partially embedded manner to form a colloidal crystal.