CELL CARRIER, ASSOCIATED METHODS FOR MAKING CELL CARRIER AND CULTURING CELLS USING THE SAME
A carrier for growing adherent cells is provided, wherein the carrier comprises one or more outer surfaces; and one or more structured indentations on one or more of the outer surfaces, wherein the carrier has a length at least about 0.2 mm, a width at least about 0.2 mm, and a height in a range from about 0.05 mm to 1.2 mm and each of the structured indentations has a major axis in a range from about 0.1 mm to 0.5 mm, a minor axis in a range from about 0.1 mm to 0.5 mm and a depth in a range from about 0.025 mm to about 0.5 mm. The carrier may comprise a single indentation or ‘cup’ like structure, or may comprise a plurality of indentations. A method of making the carrier, and culturing stromal cells using the same carrier are also provided.
1 . A carrier for growing adherent cells, comprising:
one or more outer surfaces; and
one or more structured indentations on one or more of the outer surfaces, wherein the carrier has a length at least about 0.2 mm, a width at least about 0.2 mm, and a height in a range from about 0.05 mm to 1.2 mm and each of the structured indentations has a major axis in a range from about 0.1 mm to 0.5 mm, a minor axis in a range from about 0.1 mm to 0.5 mm and a depth in a range from about 0.025 mm to about 0.5 mm.
2 . The carrier of claim 1 , wherein the carrier comprises two outer surfaces for growing adherent cells.
3 . The carrier of claim 1 , wherein each of the structured indentations has a cross sectional profile of polygonal, circular, or elliptical shape.
4 . The carrier of claim 1 , wherein each of the structured indentation has an aspect ratio in a range from about 0.1 to about 1.5.
5 . The carrier of claim 1 , wherein the carrier is made of a glass, polymer, ceramic, metal or combination thereof.
6 . The carrier of claim 1 , wherein the carrier is made of dextran, silicone, polyester, polycarbonate, polyamide, polyurethane, olefin polymer, or polyacrylate polymer.
7 . The carrier of claim 1 , wherein the carrier is made of polystyrene.
8 . The carrier of claim 1 , wherein each of the one or more surfaces is modified by a chemical functionalization, applying a coating, corona discharge treatment, or plasma treatment to form a modified surface.
9 . The carrier of claim 8 , wherein the modified surface is a cytophilic surface.
10 . The carrier of claim 8 , wherein the coating is made of thermoresponsive polymer, pH responsive polymer, or combination thereof.
11 . The carrier of claim 8 , wherein the coating is made of collagen, vitronectin, fibronectin, or laminin.
12 . The carrier of claim 1 , having a perimeter that is triangular, rectangular, square, pentagonal, hexagonal, circular, or elliptical.
13 . A cell culture system comprising one or more of the carriers of claim 1 .
14 . The cell culture system of claim 13 is a bioreactor.
15 . The cell culture system of claim 14 , wherein the bioreactor comprises fluid having a forced convective fluid motion.
16 . The cell culture system of claim 14 , wherein the bioreactor is a stirred tank bioreactor, or a reactor with rocking or rolling motion.
17 . A kit for culturing cells, comprising a disposable housing pre-loaded with the carrier of claim 1 .
18 . The kit of claim 17 , wherein the disposable housing is a bag, a flask, a tube, a petridish, or a bottle.
19 . A carrier for growing adherent cells, comprising:
one or more outer surfaces; and
a single structured indentation on at least one surface, wherein the single structured indentation has a length of at least about 1 mm, a width of at least about 1 mm, a height in a range from about 1 mm to 10 mm, and a wall-thickness of the carrier in a range from about 0.05 mm to 2 mm.
20 . A method of culturing adherent cells, comprising:
providing a carrier for growing the cells, comprising one or more outer surfaces; and one or more structured indentations on one or more of the outer surfaces, wherein the carrier has a length at least about 0.2 mm, a width at least about 0.2 mm, and a height in a range from about 0.05 mm to 1.2 mm and each of the structured indentations has a major axis in a range from about 0.1 mm to 0.5 mm, minor axis in a range from about 0.1 mm to 0.5 mm and depth in a range from about 0.025 mm to about 0.5 mm, and
growing the cells on the carrier.
21 . The method of claim 20 , wherein the cells are released after growing on the carrier using a mechanical method, a chelant, an enzyme, a thermoresponsive coating, a pH responsive coating, or a combination thereof.
22 . The method of claim 20 , wherein the adherent cells are mesenchymal stromal cells.
23 . The method of claim 22 , wherein the adherent cells are human mesenchymal stromal cells.
24 . A method of making a carrier for growing cells, comprising:
a) providing a flat polymer film;
b) forming on the flat polymer film, on one or more sides, one or more structured indentations;
c) imparting a surface treatment to the film comprising one or more of corona discharge treatment, gas plasma treatment, chemical functionalization or coating; and
d) cutting the treated polymer film into a plurality of portions.
25 . A method of making a carrier for growing cells, comprising:
a) providing one or more flat polymer films;
b) forming on the flat polymer film, on one or more sides, one or more structured indentations;
c) cutting the polymer films into a plurality of portions; and
d) imparting a treatment to the portions comprising one or more of corona discharge treatment, gas plasma treatment, chemical functionalization or coating.
26 . The method of claim 25 wherein the carrier is coated with a thermoresponsive agent, a pH responsive agent, or a combination thereof.
27 . The method of claim 25 wherein the carrier is coated with collagen, vitronectin, fibronectin, or laminin.
28 . The method of claim 25 wherein the carrier is cytophilic.
29 . A method of making a carrier for growing cells, comprising:
a) providing two flat polymer films;
b) forming on the two flat polymer films, a plurality of structured indentations on at least one surface of each of the two films;
c) laminating the two polymer films together so that at least two outwardly facing surfaces comprise a plurality of the structured indentations;
d) cutting the laminated polymer film into a plurality of portions; and
e) imparting a treatment to the portions comprising one or more of corona discharge treatment, gas plasma treatment, chemical functionalization or coating.