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"Food Safety" Research Results Published: Large-Scale Production of Chicken Skeletal Muscle Cells Using 3D Microcarrier Systems

"Food Safety" Research Results Published: Large-Scale Production of Chicken Skeletal Muscle Cells Using 3D Microcarrier Systems

  • Categories:Company News
  • Author:CytoNiche
  • Origin:CytoNiche
  • Time of issue:2024-02-28
  • Views:312

(Summary description)Using 3D TableTrix® microcarriers for suspended culture of myoblasts significantly enhances cell culture efficiency, laying the groundwork for the industrial production of cultured meat.

"Food Safety" Research Results Published: Large-Scale Production of Chicken Skeletal Muscle Cells Using 3D Microcarrier Systems

(Summary description)Using 3D TableTrix® microcarriers for suspended culture of myoblasts significantly enhances cell culture efficiency, laying the groundwork for the industrial production of cultured meat.

  • Categories:Company News
  • Author:CytoNiche
  • Origin:CytoNiche
  • Time of issue:2024-02-28
  • Views:312
Information

Preface

Recently, Senior Engineer Li Shilei from the China Meat Food Comprehensive Research Center and Researcher Liang Jun from Tianjin University jointly published a research article in "Food Science" [EI Indexed]: Large-Scale Production of Chicken Skeletal Muscle Cells Using Microcarrier Systems.

Original Article Link: 

https://www.spkx.net.cn/CN/10.7506/spkx1002-6630-20230330-310

 

Research Background

Cell-cultured meat, a novel type of meat product produced by controlling the rapid proliferation, directional differentiation, and collection and processing of animal cells through in vitro cultivation, is considered a potential solution for alleviating environmental pollution, sustainable development, and addressing global public health and animal welfare issues in the future. The production of cell-cultured meat requires large-scale in vitro cultivation of cells. However, the relatively large surface space required for cell adhesion culture limits the industrial production scale of cell-cultured meat. Therefore, to improve the efficiency of large-scale cell cultivation, researchers have focused on the development of microcarriers suitable for 3D cell culture and the development of bioreactor-related cultivation processes. By continuously improving the specific surface area utilization and biocompatibility of microcarriers, the comprehensive cultivation efficiency of cells is enhanced, and the required space volume for cell cultivation is continuously reduced.

Microcarriers are widely researched nowadays, with their practicality mainly reflected in their mechanical properties, biocompatibility, settling coefficient, degradability, and susceptibility to physical and chemical stimuli. In large-scale cell cultivation, optimizing microcarrier-based cell culture conditions is crucial for their widespread application. A common method is to construct a cellular microecosystem using microcarriers, bioreactors, cells, and culture media. Rotating flask reaction vessels are widely used for cell cultivation because they can provide a uniform culture environment. However, cells are also influenced by various external factors during the cultivation process, such as culture conditions, flask rotation speed, and microcarrier type. So far, there have been few reports on the large-scale cultivation of muscle cells for cell-cultured meat production.

This study compared C carriers and 3D TableTrix® microcarriers and selected the microcarriers more suitable for large-scale cultivation of muscle cells. Taking 3D TableTrix® microcarriers as the research object, the important parameters of cell three-dimensional cultivation were optimized. Larger-scale cell cultivation was achieved through bead-to-bead transfer, and the microcarriers with cells were cryopreserved and resuscitated. The proliferation status and morphological differences between microcarriers with cells and thawed cells were compared and observed, providing a theoretical basis and exploratory practice for large-scale cell cultivation and laying the foundation for the industrial production of cell-cultured meat.

 

Research Findings

This study found that using 3D TableTrix® microcarriers for suspension culture of muscle cells in rotating flasks significantly improved cell culture efficiency after optimizing the conditions. The 3D TableTrix® microcarriers with cells can also be cryopreserved and resuscitated for continued use, showing promising application prospects in the industrial-scale production of cell-cultured meat.

 

Research Design

Figure 1: Overview of Research Design

The schematic diagram of the research design is shown in Figure 1. Cells and 3D TableTrix® microcarriers are mixed and added to the bioreactor for cultivation. By optimizing the cell culture process, the efficiency and vitality of cell culture are improved, ultimately applying them to the industrial production of cell-cultured meat.

 

Research Content

Figure 2: Visual and Microscopic Images of 3D TableTrix® Microcarriers-Tablet (a) and C Carriers-Powder (b)

Figure 3: Particle Size Distribution of Microcarriers

Figure 4: Cell Growth Curve (a) and Differential Analysis (b)

Figure 5: Bright-field Image of Dissolved 3D TableTrix® Microcarriers

From Figures 2 to 5, it can be observed that 3D TableTrix® microcarriers are a novel type of porous gelatin microcarriers. These microcarriers are provided in the form of fixed-weight tablets, with a thickness of 1.57±0.01 mm and a diameter of 8.01±0.05 mm. On the other hand, C carriers are positively charged, non-porous polystyrene microcarriers provided in powder form. The average particle size of C carriers is significantly higher at 265 μm compared to the average particle size of 195 μm for 3D TableTrix® microcarriers. Moreover, cell proliferation efficiency on 3D TableTrix® microcarriers is significantly higher than on C carriers.

Figure 6: Impact of Different Stirring Methods on Cell Suspension Culture in Flasks

Figure 7: Impact of Different Cell Seeding Densities on Cell Culture Results

Figure 8: Impact of Different Microcarrier Mass Concentrations on Cell Culture Results

Figure 9: Differential Analysis of Cell Count at Different Rotation Speeds (a) and Growth Curve (b)

Figure 10: Differential Analysis of Cell Growth under Different Feeding Methods (a) and Growth Curve (b)

Figure 11: Impact of Different Serum Concentrations on Cell Culture Results

Figure 12: Impact of Bead-to-Bead Transfer on Cell Suspension Culture in Flasks

Figure 13: Glucose Metabolism of Cells in Flask Culture

Figure 14: Growth Curve of Cells in 2L Flask Culture

Figure 15: Effect of Microtissue Cryopreservation on Cells

In Figures 6-15, the impact of factors such as optimized initial stirring conditions, cell seeding density, microcarrier density, stirring speed, different feeding methods, serum concentration, bead-to-bead transfer, etc., on large-scale cell culture is investigated. Using single-factor variable methods, important parameters for three-dimensional cell culture are optimized and determined. Larger scale cell culture is achieved through bead-to-bead transfer, and the proliferation and morphology differences between microcarriers containing cells and thawed cells are observed and compared.

 

Research Conclusion

In this study, the 3D TableTrix® microcarrier is shown to be a dispersible and soluble porous microcarrier, with simple handling and the ability to eliminate the enzymatic dissociation process during cell passaging through bead-to-bead transfer, greatly simplifying the expansion process of myoblast cells.

Furthermore, cells can be cryopreserved on 3D TableTrix® microcarriers, and after thawing, they remain highly viable and retain their 3D macrostructure. They can serve as seed cells for subsequent batch culture through bead-to-bead transfer or as implantable carriers for cell culture. By optimizing various conditions for cell culture using 3D TableTrix® microcarriers, cell culture efficiency has been improved, and continuous culture in large-volume spinner flasks is achievable.

 

Author Information

First Author: Liu Yisen

Research focus: Large-scale cultivation of cell-cultured meat.

Corresponding Author: Li Shilei

Senior Engineer at the China Meat Food Comprehensive Research Center, specializing in research and development of cell-cultured meat and food safety. He has led or participated in several national key R&D programs, strategic consulting projects of the Chinese Academy of Engineering, Beijing Science and Technology Plan projects, Beijing Postdoctoral Foundation projects, etc.

Corresponding Author: Liang Kongjun

Researcher at Tianjin University of Science and Technology, focusing on the research of bio-based nano self-assembly materials. He has led or participated in several national key R&D programs, National Natural Science Foundation of China projects, etc.

 

Research Technical Support

 

More Biomimetic: Comprising tens of thousands of elastic three-dimensional porous microcarriers, with a porosity >90% and controllable particle sizes ranging from 50-500μm, with uniformity ≤100μm, creating a truly biomimetic 3D cell culture environment.

Full Qualification: Holds two Chinese CDE pharmaceutical excipient qualifications, with registration numbers F20200000496 and F20210000003; and three US FDA drug substance and pharmaceutical excipient qualifications, with registration numbers 037798 & 035481 and 29721.

Easy Harvesting: Incorporates specific degradation technology to achieve gentle and non-destructive cell harvesting.

Enhanced Safety: Supported by authoritative institutions' reports on residual cleavage detection, cell toxicity, pyrogenicity, genotoxicity, in vivo immunotoxicology assessments, as well as reports on hemolysis, subcutaneous injection local irritation, systemic hypersensitivity, and intraperitoneal injection toxicity.

Easy Scaling Up: Through 3D cultivation methods, combined with CytoNiche's 3D cell smart manufacturing platform, achieving fully automated closed-loop large-scale cell culture, enabling harvesting of billions of cells. Customizable microcarriers are available to meet various cell culture needs.



[About CytoNiche]

CytoNiche was established in 2018, led by Professor Yanan Du's research team from the School of Medicine at Tsinghua University, with Tsinghua University as a shareholder. The core technology originated from the transformation of scientific achievements at Tsinghua University and was recognized as a leading technology in "Science and Technology Innovation in China" by the China Association for Science and Technology. As a national-level high-tech enterprise, a national-level specialized and new technology "Little Giant" enterprise, a potential unicorn enterprise, it has also received key research and development special support from the Ministry of Science and Technology.

As an expert in high-quality three-dimensional cell manufacturing, CytoNiche provides a one-stop customized solution for cell scale-up based on 3D microcarriers. The company has built an original 3D cell smart manufacturing platform, achieving large-scale, automated, intelligent, and closed-cell drug and derivative production preparation. This helps global customers establish the most advanced cell drug production lines. After pioneering the production process pipeline for "billion-level" stem cells, the company is accelerating towards "hundred billion-level," dedicating efforts to empower the cell and gene therapy industry with 3D cell scale-up smart manufacturing technology to benefit more patients.

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Copyright: Beijing CytoNiche Biotechnology Co., Ltd.
Copyright: Beijing CytoNiche Biotechnology Co., Ltd.