Method for cell culture
View Patent ↗A method for stem or progenitor cell culture. More precisely, the invention relates to a method for cell culture using one or more IαI (inter-alpha trypsin inhibitor or inter-alpha inhibitor) protein(s) or part(s) thereof as a component in a cell culture media or a coating on a cell culture surface material. Furthermore the invention relates to a cell culture media and a cell culture coating/matrix provided with one or more IαI proteins(s) or part(s) thereof.
1. A cell culture surface comprising a coating comprising one or more inter alpha trypsin inhibitor (IαI) proteins in a concentration of 2 μg/ml-100 μg/ml,
wherein the one or more proteins are selected from the group consisting of an IαI protein comprising heavy chain 1 (HC1), heavy chain 2 (HC2), and bikunin (B) (IαI -HC1, HC2, B), an isolated HC2, and any other IαI protein comprising heavy chain 2 (IαI -HC2),
wherein the cell culture surface supplements a cell culture medium that is partially or completely chemically defined, and
wherein the cell culture medium is whole serum free, serum component free and feeder cell free.
2. The cell culture surface of claim 1 , wherein the concentration of the one or more proteins is 10 μg/mL-50 μg/mL.
3. The cell culture surface of claim 1 , wherein the one or more IαI proteins are un-complexed proteins isolated from serum or a serum fraction, produced as a recombinant protein, synthesized chemically, or a combination thereof.
4. The cell culture surface of claim 1 , wherein the one or more IαI proteins comprise the IαI- HC2 protein.
5. A method of making the cell culture surface of claim 1 , the method adding the one or more IαI proteins onto the uncoated cell culture surface and allowing the coating to form on the uncoated cell culture surface.
6. The method of claim 5 , wherein the coating formation takes place at 4° C. overnight.
7. The method of claim 5 , wherein the coating formation takes place at room temperature or 37° C. in 1-2 hours.
8. A cell culture surface comprising a coating comprising an IαI -HC1, HC2, B protein in a concentration of 2 μg/ml-200 μg/ml,
wherein the cell culture surface supplements a cell culture medium that is partially or completely chemically defined, and
wherein the cell culture medium is whole serum free, serum component free and feeder cell free.
9. The cell culture surface of claim 8 , wherein the concentration of the IαI -HC1, HC2, B protein is 10 μg/mL-50 μg/mL.
10. The cell culture surface of claim 8 , wherein the IαI -HC1, HC2, B protein is an un-complexed protein isolated from serum or a serum fraction, produced as a recombinant protein, synthesized chemically, or a combination thereof.
11. A method of making the cell culture surface of claim 8 , the method comprising coating an uncoated cell culture surface with the IαI -HC1, HC2, B by adding the IαI -HC1, HC2, B protein onto the uncoated cell culture surface and allowing the coating to form on the uncoated cell culture surface.
12. The method of claim 11 , wherein the coating formation takes place at 4° C. overnight.
13. The method of claim 11 , wherein the coating formation takes place at room temperature or 37° C. in 1-2 hours.
14. A cell culture surface comprising a coating comprising an isolated HC2 at a concentration of 2 μg/ml-200 μg/ml,
wherein the cell culture surface supplements a cell culture medium that is partially or completely chemically defined, and
wherein the cell culture medium is whole serum free, serum component free and feeder cell free.
15. The cell culture surface of claim 14 , wherein the concentration of the isolated HC2 protein is 10 μg/mL-50 μg/mL.
16. The cell culture surface of claim 14 , wherein the isolated HC2 is an un-complexed protein isolated from serum or a serum fraction, produced as a recombinant protein, synthesized chemically, or a combination thereof.
17. A method of making the cell culture surface of claim 14 , the method comprising coating an uncoated cell culture surface with the IαI-HC2 by adding the isolated HC2 onto the uncoated cell culture surface and allowing the coating to form on the uncoated cell culture surface.
18. The method of claim 17 , wherein the coating formation takes place at 4° C. overnight.
19. The method of claim 17 , wherein the coating formation takes place at room temperature or 37° C. in 1-2 hours.