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Method for preparing cell culture meat from high-activity muscle stem cells

A cell culture and stem cell technology, applied in the field of stem cell applications, can solve problems such as lack of culture and differentiation, achieve efficient and rapid growth, promote the formation of muscle fibers, and rationally arrange time and space advantages

Active Publication Date: 2022-03-04
SHANDONG FIRST MEDICAL UNIV & SHANDONG ACADEMY OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, how to achieve high-efficiency culture and differentiation lacks corresponding technologies, especially the selection of highly active muscle stem cells at the beginning, as well as the time and space coordination of the two processes of rapid and large-scale expansion of cells in vitro and myogenic differentiation.

Method used

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  • Method for preparing cell culture meat from high-activity muscle stem cells
  • Method for preparing cell culture meat from high-activity muscle stem cells
  • Method for preparing cell culture meat from high-activity muscle stem cells

Examples

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Effect test

Embodiment 1

[0031] A method for producing cell cultured meat by utilizing dog's highly active muscle stem cells, the specific steps are as follows:

[0032] (1) Screening to obtain muscle stem cells with high functional activity: Primary muscle stem cells from the hind limbs of experimental dogs (Golden Retriever, 2 years old) were initially cultured and expanded to about 5×10 4 After the number of cells, they were separated and screened according to their ALDH1 activity, and about 15% of the cells with the highest ALDH1 activity were isolated (~1x10 4 cell);

[0033](2) Efficiently obtain hydrogel monolayer cells of highly active muscle stem cells: when ALDH1 highly expressed muscle stem cells are cultured and expanded to the fourth generation, they are introduced into the hydrogel thin layer structure formed by collagen (~2mm Thickness), use this 3D cell culture substrate for large-scale cell expansion. A polypeptide fragment of 0.5 mg / mL Jagged1 protein is combined in the hydrogel to...

Embodiment 2

[0038] A method for producing cell-cultured meat using highly active muscle stem cells of cattle, the specific steps are as follows:

[0039] (1) Muscle stem cells with high functional activity were screened: the primary muscle stem cells from the legs of pasture-raised cattle (Black Angus) were initially cultured and expanded to about 5x10 4 After the number of cells, according to the activity of ALDH1, isolate and screen, and isolate about 30% of the cells with the highest ALDH1 activity (~1.5x10 4 cell);

[0040] (2) Efficiently obtain hydrogel monolayer cells with high activity of muscle stem cells: when ALDH1 highly expressed muscle stem cells are cultured and expanded to the sixth passage, they are introduced into the hydrogel thin layer structure formed by collagen (~2mm Thickness), use this 3D cell culture substrate for large-scale cell expansion. 0.5mg / mL polypeptide fragments of Jagged1 protein are bound in the hydrogel. At the same time, Follistatin 344 (150ng / mL...

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Abstract

The invention belongs to the field of stem cell application, and particularly relates to a method for preparing animal cell culture meat from high-activity muscle stem cells. A polypeptide fragment of Jagged1 is combined in the hydrogel cell scaffold in advance, and a Notch signal channel of the stem cells is effectively activated, so that the characteristics (dryness and growth differentiation potential) of the stem cells are maintained. The added Follistatin can inhibit the activity of myostatin, so that the volume, growth and differentiation of muscle cells are more potential, and the differentiated muscle fibers are thicker and stronger. And the more mature and robust muscle fibers can enhance the texture and taste of the cell culture meat and generate more special nutritional factors of the mature muscle cells. Meanwhile, the hydrogel thin layers on which the muscle cells grow are overlaid in multiple layers to form the three-dimensional muscle tissue, and differentiation of cell myogenic fibers under the low-serum condition is continued, so that the growth and differentiation time and space advantages of the muscle stem cells are arranged more reasonably.

Description

technical field [0001] The invention belongs to the field of stem cell application, in particular to a method for preparing cell cultured meat by utilizing highly active muscle stem cells. Background technique [0002] Artificial meat, that is, cell-based meat (CBM), has received extensive attention from the global public opinion and market in recent years. Global meat production and consumption continues to surge due to population growth, growing personal and socioeconomic needs. In 2012, the Food and Agriculture Organization of the United Nations (FAO) predicted that by 2050, global meat demand will reach 455 million tons (a 76% increase from 2005). Growing global demand for meat is a growing concern due to the complexity of current large-scale livestock production methods and public health issues, as well as the greenhouse effect, environmental degradation and animal welfare associated with meat production and negative consequences. The large-scale development of anima...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077C12N13/00
CPCC12N5/0659C12N13/00C12N5/0658C12N2513/00C12N2533/54C12N2501/42C12N2501/31C12N2529/00C12N2500/80
Inventor 牟晓东牟艳玲甘元善戴晓宇李栋姚庆强刘蕾
Owner SHANDONG FIRST MEDICAL UNIV & SHANDONG ACADEMY OF MEDICAL SCI
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