Cell culture method using laminin fragment-containing medium

Patent metadata

Field	Value
Publication number	US-2018334656-A1
Application number	US-201615771329-A
Country	US
Kind code	A1
Filing date	Nov 8, 2016
Priority date	Nov 10, 2015
Publication date	Nov 22, 2018
Grant date	—

Abstract

Provided is a cell culture method comprising the step of culturing cells using a medium containing a laminin fragment having integrin binding activity, the method not comprising the step of coating a culture vessel with a laminin or a laminin fragment before seeding the cells in the culture vessel. The cell culture method of the present invention uses a smaller amount of a laminin fragment and still achieves a comparable culture efficiency as compared with the conventional cell culture method that uses a culture vessel precoated with a laminin or a laminin fragment.

First claim

1 . A cell culture method comprising:
culturing cells in a medium containing a laminin fragment having integrin binding activity, the cells being mammalian stem cells or cells differentiated from the mammalian stem cells, the laminin fragment being present in the medium at a concentration of 0.03 μg to 0.5 μg of the laminin fragment per cm 2 of culture surface area of a culture vessel, the method not comprising the step of coating the culture vessel with a laminin or a laminin fragment before seeding the cells in the culture vessel.

2 - 11 . (canceled)

12 . The cell culture method according to claim 1 , wherein the cells are seeded and cultured in a culture vessel, which is not precoated with a laminin or a laminin fragment.

13 . The cell culture method according to claim 1 , wherein the laminin fragment having integrin binding activity is a laminin E8 fragment.

14 . The cell culture method according to claim 1 , wherein the laminin fragment having integrin binding activity is present in the medium at a concentration of 0.1 μg to 0.25 μg of the laminin fragment per cm 2 of culture surface area of the culture vessel.

15 . The cell culture method according to claim 1 , wherein the integrin is integrin α6β1, integrin α6β4, integrin α3β1 or integrin α7β1.

16 . The cell culture method according to claim 1 , wherein the cells are pluripotent stem cells.

17 . The cell culture method according to claim 16 , wherein the pluripotent stem cells are induced pluripotent stem cells (iPS cells). Filing Date: Herewith

18 . The cell culture method according to claim 16 , wherein the pluripotent stem cells are embryonic stem cells (ES cells).

19 . A cell culture medium for mammalian stem cells or cells differentiated from the mammalian stem cells, comprising a laminin E8 fragment having integrin binding activity.

20 . The cell culture medium according to claim 19 , wherein the laminin E8 fragment having integrin binding activity is present in said cell culture medium at a concentration of 0.15 μg/mL to 2.5 μg/mL.

21 . The cell culture method according to claim 12 , wherein the laminin fragment having integrin binding activity is a laminin E8 fragment.

22 . The cell culture method according to claim 12 , wherein the integrin is integrin α6β1, integrin α6β4, integrin α3β1 or integrin α7β1.

Assignees

• Univ Kyoto

Inventors

• Suemori Hirofumi
• Miyazaki Takamichi

Classifications

• C12N2500/30

  Organic components · CPC title

• C12N5/0696 Primary

  Artificially induced pluripotent stem cells, e.g. iPS · CPC title

• C12N5/0606 Primary

  Pluripotent embryonic cells, e.g. embryonic stem cells [ES] (embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696) · CPC title

• C12N2533/52

  Fibronectin; Laminin · CPC title

• C12N2501/585

  Integrins · CPC title