Cell culture sheet and large-capacity cell culture device including same

Patent metadata

Field	Value
Publication number	US-2022340852-A1
Application number	US-202017640970-A
Country	US
Kind code	A1
Filing date	Sep 7, 2020
Priority date	Sep 6, 2019
Publication date	Oct 27, 2022
Grant date	—

Abstract

Provided is a cell culture sheet. A cell culture sheet according to an embodiment of the present invention includes: a fiber web which has a 3-dimensional network structure formed through the accumulation of support fibers having an average diameter of at most 1.5 μm, and has a basis weight of 1 to 15 g/m2; and a functional coating layer which is coated on the support fibers exposed on at least one surface of the fiber web, and has a function of promoting one or more of the adhesion, movement, proliferation, and differentiation of cells. Accordingly, cell adhesion is improved due to the large specific surface area and the surface morphology suitable for cells, and the adhered cells are stably supported. Moreover, the cells can be cultured at high density at a high culture efficiency, and can be cultured and recovered without agglomeration caused by forming a thin film.

First claim

1 . A cell culture sheet, comprising:
a fiber web which has a 3-dimensional network structure formed through the accumulation of support fibers having an average diameter of at most 1.5 μm, and has a basis weight of 1 to 15 g/m 2 ; and a functional coating layer which is coated on the support fibers exposed on at least one surface of the fiber web, and has a function of promoting one or more of the adhesion, movement, proliferation and differentiation of cells.

2 . The cell culture sheet of claim 1 , wherein the cell culture sheet is for culturing stem cells.

3 . The cell culture sheet of claim 2 , wherein the stem cells are at least one selected from the group consisting of human induced pluripotent stem cells (hiPSC), human cardiac stem cells (hCSC), mesenchymal stem cells (MSC), murine embryonic stem cells (mESCs) and osteoblasts.

4 . The cell culture sheet of claim 1 , wherein the support fibers comprise polyvinylidene fluoride (PVDF).

5 . The cell culture sheet of claim 1 , wherein the center line average roughness (Ra) of one surface of the fiber web on which the functional coating layer is formed is 0.15 to 1.0 μm, and the developed surface area ratio (Sdr) is 1.3 to 3.0.

6 . The cell culture sheet of claim 1 , wherein the cell culture sheet is for culturing stem cells, the average diameter of the support fibers is 200 to 600 nm, the center line average roughness (Ra) of one surface of the fiber web on which the functional coating layer is formed is 0.15 to 0.6 μm, and the developed surface area ratio (Sdr) is 1.3 to 2.3.

7 . The cell culture sheet of claim 1 , further comprising a support film fixed to one surface of the fiber web.

8 . The cell culture sheet of claim 7 , further comprising a silicone-based adhesive layer between the support film and the fiber web.

9 . The cell culture sheet of claim 1 , wherein the number of cells recovered after culturing under Culture Condition 1 below is 9 times or more per unit area (cm 2 ) compared to the number of seeded cells:
[Culture Condition 1] 60 cell culture sheets at 25 cm×25 cm in width and length per sheet are fixed inside a housing by spacing apart such that the top and bottom interval between the cell culture sheets is 1 mm, and after a medium mixed with stem cells is injected into the housing, the housing is sealed from the outside air and cultured at 37° C. for 4 days.

10 . The cell culture sheet of claim 1 , wherein the number of cells recovered after culturing under Culture Condition 2 below is 25 times or more per unit area (cm 2 ) compared to the number of seeded cells:
[Culture Condition 2] 100 cell culture sheets at 25 cm×25 cm in width and length per sheet are fixed inside a housing by spacing apart such that the top and bottom interval between the cell culture sheets is 1 mm, and after a medium mixed with stem cells is injected into the housing, the housing is sealed from the outside air and cultured at 37° C. for 5 days, and after 24 hours after seeding the cells, it is cultured by replacing once with the same medium that does not include stem cells.

11 . The cell culture sheet of claim 9 , wherein the average diameter of the stem cells recovered after culturing under Culture Condition 1 is smaller by 15% or more than the average diameter of the seeded stem cells.

12 . The cell culture sheet of claim 11 , wherein the average diameter of the stem cells recovered after the culture is 18 μm or less.

13 . The cell culture sheet of claim 1 , wherein the functional coating layer comprises a fusion protein between a functional peptide and an adhesion protein.

14 . A large-capacity cell culture device, comprising:
a housing; and the cell culture sheet according to claim 1 , which is provided in a plurality of sheets inside the housing and is arranged in multiple stages at a predetermined interval along one direction.

15 . A large-capacity cell culture system, comprising:
the large-capacity cell culture device according to claim 14 ; a medium supply device for supplying a medium necessary for cell culture to one side of the large-capacity cell culture device; and a pump for circulating the medium.

16 . The cell culture sheet of claim 10 , wherein the average diameter of the stem cells recovered after culturing under Culture Condition 2 is smaller by 15% or more than the average diameter of the seeded stem cells.

17 . The cell culture sheet of claim 16 , wherein the average diameter of the stem cells recovered after the culture is 18 μm or less.

Assignees

• Amolifescience Co Ltd

Inventors

• Jang Seon Ho
• Seo In Yong
• Han Kyoung Ku
• Seo Dong Sik
• Park Hee Sung
• Lee Su Yeon
• Lee Hyo Jung
• Kim Ji Young
• Kim Chan
• Lee Seoung Hoon

Classifications

• C12N5/0062 Primary

  General methods for three-dimensional culture · CPC title

• C12N5/0068 Primary

  General culture methods using substrates (for specific animal cell type C12N5/06) · CPC title

• C12N5/0662

  Stem cells · CPC title

• C12N2533/30

  Synthetic polymers · CPC title

• C12N2523/00

  Culture process characterised by temperature · CPC title