Approaching 3d cell culture

  • The three-dimensional cell culture is mainly based on the 3d cell culture scaffold model, which can better simulate the growth environment of the cells in the body.

     

    Definition:

    Three-dimensional cell culture technology, ie 3d cell culture techniques, means that there will be the carrier of 3d structurally different materials is co-cultured with various kinds of cells in vitro, so that the cells can migrate and grow in the 3D spatial structure of the vector to form a 3d cell-carrier complex.

    Application:

    3d cell culture applications are particularly important than normal cell culture. Ordinary cell culture gradually loses its original traits due to the proliferation of cells in an environment that changes in vitro, often inconsistent with the in vivo conditions, and animal experiments are completely carried out in vivo, but due to various factors in the body and the internal and external environment It becomes complicated by influence, it is difficult to study a single process, and it is difficult to study an intermediate process. 3d cell culture technology is a technology between monolayer cell culture and animal experiments. It can not only simulate the internal environment to the greatest extent, but also demonstrate the advantages of cell culture intuitiveness and conditional controllability.

    1. Tumor biology: experimental treatment of tumors, invasiveness of tumors, mechanisms of metastasis and central necrosis, tumor angiogenesis and nutrient supply, gene expression in vivo, test drug inhibition of tumor growth and inhibition to adjacent tissues etc.
    2. Cartilage and bone tissue: Mature chondrocytes and stem cells are widely used in 3d cell cultureto regenerate damaged cartilage, bone, ligament, tendon and knee meniscus.
    3. Circulatory system and heart: The timely generation of vascular networks in mature tissues is an important topic in tissue engineering; in the therapeutic field, it is also concerned how to purposefully change blood vessel formation to inhibit tumor progression.
    4. nervous system: culture a single neuron into a multi-cell aggregate, hippocampus live specimens, test neuronal potential, neural stem cell culture treatment of Alzheimer's disease, Parkinson's disease.

     

    3d cell culture research frontier:

    1. 3d cell tissue plus tensile culture model TissueTrain tensile stress stimulation 3d hydrogel stent cell tissue culture system

    Functional highlights of the 3d cell tissue augmentation model: true 3D culture - the system is coated with a variety of surfaces (Amino, Collagen (Type I or IV), Elastin, ProNectin (RGD), Laminin (YIGSR)) The collagen hydrogel is an extracellular matrix scaffold in the study of biomaterial scaffolds. Compared with traditional nanofiber scaffolds and porous scaffolds, the hydrogel scaffold cross-linking network contains a large amount of water, which can supply cell nutrients well. It is also possible to cross-link biologically active factors to regulate cell growth and differentiation. Therefore, hydrogel scaffolds can better simulate the tissue-like physical and spatial structures required for cell growth, and have high plasticity, relatively simple manufacturing process, and convenient clinical application. Because collagen is the most abundant protein in the human body (about 25%), it is the most common protein in the extracellular matrix. On the collagen fiber, there are amino acid sequences such as arginine-glycine aspartate, which can be cells. Recognition and attachment by surface integrin. Moreover, collagen itself is a natural material with small immunorejection reaction, and its cross-linking process does not require the introduction of other chemical agents, and can be self-crosslinked to form a gel 3d scaffold, and its biocompatibility is more prominent.

    Therefore, collagen hydrogels have received widespread attention. In the past few decades, tissue engineering experts have worked to develop materials that better simulate 3d culture analogs to overcome the shortcomings of two-dimensional cell culture. To this end, cells are seeded in microporous scaffolds, nanofiber scaffolds, and hydrogel scaffolds for culture. The microporous scaffold is easy to use, but its pore size is much larger than the average cell diameter, so it is actually equivalent to two-dimensional culture. The nanofiber scaffold uses a fibrous extracellular matrix protein to better simulate the 3d structure, but its mechanical properties are difficult to meet the requirements of use. The hydrogel scaffold encapsulates cells in a liquid state, forming a cross-linking network in a solid state, allowing a large number of cells to be dispersed and adhered therein, so that the transplanted cells can contact the matrix, which is equivalent to the true 3d culture. Moreover, the collagen gel is a hydrogel, and the nutrients can enter and exit the gel network freely, so that the cells dispersed in the network can be nutritious, so the collagen hydrogel has good hydrophilicity and cell compatibility. In addition, liquid collagen is easy to add various growth factors and plays an important role in cell growth and differentiation.

     

     

    1) 3d cell stretch stress loading stimulation: static or periodic stress stimulation of cells grown in the 3d state;

    2) 3d cell culture: 3d cell culture can be performed using 3d tissue culture mold and 3d cell culture plate;

    3) 3d cell stress loading: uniaxial or biaxial static or periodic stress loading experiments were performed on cells grown in 3d environment by Flexcell stress loading system and arc rectangular loading platform;

    4) Dynamic simulation experiment: Various special simulation experiments can be established: heart rate simulation experiment, walking simulation experiment, running simulation experiment and other dynamic simulation experiments;

    5) Bioartificial tissue construction: bioartificial tissue up to 35 mm in length can be constructed;

    6) Observing real-time reflection under cell stress: real-time observation of the reaction of the cells in the 3d state using a microscope;

    7) A variety of matrix protein coated nylon mesh anchors can enhance the binding of cells to the mesh anchor.

     

    1. 3d cell tissue aftercing culture model 3d cell tissue pressure loading culture system model

    1) The system provides periodic or static pressure loading of various tissue, 3d cell cultures;

    2) Based on the deformation of the flexible film substrate, the force is uniform;

    3) The reaction of cells and tissues under pressure can be observed in real time;

    4) can selectively block stress loading on cells;

    5) Simultaneously have multi-channel cell pull force loading function;

    6) Up to 4 channels, 4 different programs can be run simultaneously, and multiple different pressure deformation rate comparison experiments are performed;

    7) Multiple frequencies (0.01-5 Hz), multiple amplitudes and multiple waveforms can be operated in the same program;

    8) Better control of waveforms under ultra-low or ultra-high stress;

    9) Multiple waveform types: static waveform, positive rotation waveform, cardiac waveform, triangular waveform, rectangle, and various special waveforms;

    10) Computer system accurately and intelligently regulates pressure loading cycle, size, frequency and duration.

     

     

    Typical application range:

    Detection of biochemical reactions of various tissues and cells under pressure, such as: gastric epithelial cells, intestinal epithelial cells, cartilage tissue, intervertebral disc tissue, tendon tissue, ligament tissue, and from muscle, lung (lung cells), heart, Cells isolated from blood vessels, skin, tendons, ligaments, cartilage, and bone.

     

    1. 3d cell tissue aftercing culture model 3d cell tissue stretch tensile force loading culture system model

    1) The system provides axial and circumferential stress loading on two-dimensional, 3d cells and tissues;

    2) Based on the deformation of the flexible film substrate, the force is uniform;

    3) The reaction of cells and tissues under stress can be observed in real time;

    4) can selectively block stress loading on cells;

    5) Simultaneous multi-channel cell pressure loading function;

    6) Compatible with the Flex Flow parallel slab flow chamber, which can apply fluid shear stress while pulling the cells;

    7) Up to 4 channels, 4 different programs can be run simultaneously, and multiple different tensile deformation rate comparison experiments are performed;

    8) Multiple frequencies, multiple amplitudes and multiple waveforms can be operated in the same program;

    9) Better control of waveforms under ultra-low or ultra-high stress;

    10) Multiple waveform types: static waveform, positive rotation waveform, cardiac waveform, triangular waveform, rectangle, and various special waveforms;

    11) The computer system accurately and intelligently regulates the loading period, size, frequency and duration of the stretching force.

     

    Typical application range:

    Loading analyzes the biochemical reactions of various cells under stress stimulation. For example: bone cells, lung cells, cardiomyocytes, blood cells, skin cells, tendon cells, ligament cells, chondrocytes and bone cells, kidney bladder cells, smooth muscle cells/urothelium and urothelial cells, ocular epithelial cells, small eyes Tensile tissue cells, renal tubular epithelial cells, intestinal epithelial cells, gastric epithelial cells and the like are stretched and stretched.

    Common cell models:

    1. Spontaneous cell aggregation
    2. Substrate coverage culture
    3. Rotating flask culture
    4. Microcarrier culture
    5. Preset bracket culture
    6. Rotating cell culture system
    7. Cell tensile stress loading culture
    8. Cytostatic stress loading culture
    9. Cell fluid shear stress loading culture
    10. 3d cell hydrogel scaffold culture