Cultivating method used for coronary camber tissue under composite load and bioreactor thereof
A culture method and composite load technology, applied in the fields of biomedical engineering and tissue engineering, can solve the problems of culture growth and development not meeting clinical requirements, etc.
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Embodiment 1
[0030] According to the present invention, the mechanical conditions and / or hypoxic environment of rolling load or sliding load and / or torsional shear load and / or vibration load are provided, and the method for culturing tissue with coronal curved surface (including planar shape), is carried out in a bioreactor:
[0031] It includes a cell culture incubator with hypoxia and 37-degree temperature adjustment, and perfusion conditions. Under sterile conditions, the culture is transferred to a sterilized culture room, and the culture is fixed according to the standard process of cell and tissue culture. On the bottom plate of the culture room, the peristaltic pump drives the perfusion of the culture solution to keep the culture solution fresh, and add active factors (such as insulin-like growth factor (IGF), transforming growth factor-β (TGF-β), basic fibroblast growth factor) to the culture solution. Factor (bFGF), bone morphogenetic protein (BMP), platelet-derived growth factor ...
Embodiment 2
[0042] First make the culture of the coronal surface. Chondrocytes are inoculated on the coronal collagen scaffold; at the same time, osteoblasts are inoculated on the spherical-segment-shaped cancellous bone scaffold, and then the chondrocyte-collagen complex is superimposed on the spherical-segment-shaped complex to form a culture.
[0043] The roller is a rotating body whose base line of rotation conforms to the shape of the cartilage.
[0044] The culture environment is: under the condition of 5% oxygen content, rolling load is applied first, then sliding load, then torsional load, and then vibration load, and each load is applied for 10 minutes, 3 times within 24 hours. Each load is described in detail below.
[0045] Loading of rolling load: the rolling speed of the roller is 1cm / s, the reciprocating rolling frequency is 0.1Hz, and the cartilage culture deformation is 20% at the rolling contact of the roller; the roller is made of stainless steel, and there is no silico...
Embodiment 3
[0050] The mechanical conditions of rolling load or sliding load and / or torsional shear load and / or vibration load and / or hypoxic environment provided by the present invention can be used in bioreactors with crown curved surface tissue culture. Mainly be designed for the method proposed in the present invention, as Figure 1 to Figure 6 As shown, the bioreactor includes a three-gas incubator, a culture solution perfusion system, a rolling and sliding loading mechanism, a torsion-vibration mechanism, a culture chamber 4, an upper bottom plate 10, a lower bottom plate 16, a pillar 12, and a computer system.
[0051] The upper base plate 10 and the lower base plate 16 are fixed into a frame structure through four pillars 12, and the rolling and sliding loading mechanism and the cultivation chamber 4 are fixed on the upper base plate 10 through two vertical flat-shaped brackets 9 superior.
[0052] The rolling loading mechanism is a pure rolling mechanism that makes a roller roll...
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