Temperature-sensitive cell culture composition, method for using the same, method for forming the same, and use thereof

Patent metadata

Field	Value
Publication number	US-12227759-B2
Application number	US-202017126942-A
Country	US
Kind code	B2
Filing date	Dec 18, 2020
Priority date	Dec 20, 2019
Publication date	Feb 18, 2025
Grant date	Feb 18, 2025

Abstract

A temperature-sensitive cell culture composition is provided. The temperature-sensitive cell culture composition includes a hydrogel, a cellulose, a gelatin and a collagen. Based on 1 part by weight of the collagen, a content of the hydrogel is between 0.03 parts by weight and 60 parts by weight, a content of the cellulose is between 150 parts by weight and 360 parts by weight, and a content of the gelatin is between 21 parts by weight and 12 parts by weight. In addition, a method for using the temperature-sensitive cell culture composition, a method for forming the temperature-sensitive cell culture composition, and a use of the temperature-sensitive cell culture composition are also provided.

First claim

What is claimed is:

1. A temperature-sensitive cell culture composition, comprising:
a hydrogel, wherein the hydrogel has a structure represented by Formula (I) or Formula (II) shown as follows:
A-B-BOX-B-A,  Formula (I); or

B-A-B-(BOX-BAB)n-BOX-B-A-B,  Formula (II),

wherein the polymer block A is a hydrophilic polyethylene glycol polymer, the polymer block B is a hydrophobic polyester polymer, BOX is a bifunctional group monomer of 2,2′-Bis(2-oxazoline) for coupling di-block of A-B or tri-block of B-A-B, and n is an integer greater than or equal to 0, and wherein an average molecular weight of the hydrogel is between 2000 and 6000,
wherein the polymer block A comprises polyethylene glycol (PEG) or methoxy-poly(ethylene glycol) (mPEG), and the polymer block B comprises poly(lactide-co-glycolide) (PLGA), poly(propionic-co-lactic (PPLA), poly(valeric-co-lactic) (PVLA) or poly(caproic-co-lactic) (PCLA);
a cellulose;
a gelatin; and
a collagen;
wherein based on 1 part by weight of the collagen, a content of the hydrogel is between 0.03 parts by weight and 60 parts by weight, a content of the cellulose is between 150 parts by weight and 360 parts by weight, and a content of the gelatin is between 1 part by weight and 12 parts by weight.

2. The temperature-sensitive cell culture composition as claimed in claim 1 , wherein based on 1 part by weight of the collagen, a content of the hydrogel is between 0.33 parts by weight and 33 parts by weight, a content of the cellulose is between 166 parts by weight and 333 parts by weight, and a content of the gelatin is between 3 parts by weight and 10 parts by weight.

3. The temperature-sensitive cell culture composition as claimed in claim 1 , wherein the collagen comprises atelocollagen.

4. The temperature-sensitive cell culture composition as claimed in claim 1 , wherein the cellulose comprises methyl cellulose.

5. The temperature-sensitive cell culture composition as claimed in claim 1 , further comprising a medium, the medium comprises amino acids, vitamins, serum, antibiotics or a combination thereof.

6. The temperature-sensitive cell culture composition as claimed in claim 1 , further comprising water.

7. A method for using a temperature-sensitive cell culture composition, comprising:
adding a cell to the temperature-sensitive cell culture composition as claimed in claim 1 at a first temperature; and
performing cell culture at a second temperature.

8. The method for using a temperature-sensitive cell culture composition as claimed in claim 7 , wherein the temperature-sensitive cell culture composition is a liquid at the first temperature, the temperature-sensitive cell culture composition is a gel at the second temperature.

9. The method for using a temperature-sensitive cell culture composition as claimed in claim 7 , wherein the first temperature is between 0° C. and 32° C.

10. The method for using a temperature-sensitive cell culture composition as claimed in claim 7 , wherein the second temperature is between 33° C. and 40° C.

11. The method for using a temperature-sensitive cell culture composition as claimed in claim 7 , further comprising:
after culturing cells at the second temperature, loading the temperature-sensitive cell culture composition into an appliance at the first temperature.

12. The method for using a temperature-sensitive cell culture composition as claimed in claim 11 , further comprising:
applying the temperature-sensitive cell culture composition to a subject by the appliance.

13. The method for using a temperature-sensitive cell culture composition as claimed in claim 7 , wherein the cell comprises a stem cell, an immune cell, a somatic cell or a combination thereof.

14. A method for forming a temperature-sensitive cell culture composition, comprising:
mixing a hydrogel and a cellulose at a first temperature, allowing to stand for a first time interval at a second temperature to form a gel substrate, wherein the hydrogel has a structure represented by Formula (I) or Formula (II) shown as follows:
A-B-BOX-B-A,  Formula (I); or

B-A-B-(BOX-BAB)n-BOX-B-A-B,  Formula (II),

wherein the polymer block A is a hydrophilic polyethylene glycol polymer, the polymer block B is a hydrophobic polyester polymer, BOX is a bifunctional group monomer of 2,2′-Bis(2-oxazoline) for coupling di-block of A-B or tri-block of B-A-B, and n is an integer greater than or equal to 0, and wherein an average molecular weight of the hydrogel is between 2000 and 6000,
wherein the polymer block A comprises polyethylene glycol (PEG) or methoxy-poly(ethylene glycol) (mPEG), and the polymer block B comprises poly(lactide-co-glycolide) (PLGA), poly(propionic-co-lactic (PPLA), poly(valeric-co-lactic) (PVLA) or poly(caproic-co-lactic) (PCLA; and
adding a mixture of a collagen and a gelatin to the gel substrate;
wherein based on 1 part by weight of the collagen, a content of the hydrogel is between 0.03 parts by weight and 60 parts by weight, a content of the cellulose is between 150 parts by weight and 360 parts by weight, and a content of the gelatin is between 1 part by weight and 12 parts by weight.

15. The method for forming a temperature-sensitive cell culture composition as claimed in claim 14 , wherein after the mixture of the collagen and the gelatin is added to the gel substrate, it is allowed to stand at the second temperature for a second time interval, so that the mixture of the collagen and the gelatin forms a gel layer.

16. The method for forming a temperature-sensitive cell culture composition as claimed in claim 15 , further comprising:
adding a culture medium on the gel layer, wherein the culture medium is a liquid.

Assignees

• Ind Tech Res Inst

Inventors

• Lu Chia-Jung
• Huang Jing-En
• Su Liang-Cheng
• Shen Hsin-Hsin
• Wang Yuchi
• Chao Ying-Hsueh
• Lin Li-Hsin
• Huang Hsiu-Hua

Classifications

• A61K47/42

  Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein (oligopeptides having up to five amino acids {A61K47/183}; polyamino acids A61K47/34) · CPC title

• C12N2533/40

  Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers · CPC title

• C12N2533/54

  Collagen; Gelatin · CPC title

• C12N2533/78

  Cellulose · CPC title

• C12N2523/00

  Culture process characterised by temperature · CPC title