Manufacturing method of bracket material for applying in myocardial tissue engineering
A technology of tissue engineering and scaffold materials, applied in tissue culture, bone/connective tissue cells, biochemical equipment and methods, etc., can solve problems such as unsatisfactory quality, and achieve beneficial effects on development and function
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Embodiment 1
[0006] Example 1 Preparation of a scaffold that simulates myocardial extracellular matrix
[0007] Collagen comes from pig skin and is mainly composed of types I and III. Commercially available pigskin, remove fat, sundries, etc., shred and dissolve in 0.1% acetic acid. Centrifuge at 2000 rpm to remove undissolved impurities. The collagen in the supernatant solution was precipitated with 4M NaCl. The precipitated collagen was put into a dialysis bag and dialyzed to remove NaCl. The collagen after removing NaCl was dissolved in 0.1% acetic acid again to obtain the desired collagen solution. Collagen and elastin (Sigma Company) were mixed 1:1 to obtain mixture 1, and 20% horse serum (Beijing Yuanheng Sanma Biotechnology Research Institute) and 4% chicken serum were added to 2×DMEM (Gibco BRL or Sigma Company). Embryo extract (self-made) to obtain mixture 2, mixture 1 and mixture 2 were mixed at a ratio of 1:1 to obtain mixture 3, mixture 3 was neutralized with 0.1M NaOH to o...
Embodiment 2
[0008] Example 2 Construction of tissue-engineered myocardium using a scaffold that simulates the extracellular matrix of the myocardium
[0009] Build support material by embodiment 1. Mix freshly isolated neonatal rat (day 1-2) cardiomyocytes with the scaffold material to a final concentration of 2×10 6 / ml or so. The mixed solution was added to the ring tank, and the culture solution (DMEM containing 10% horse serum) was added after 60 minutes. The mixture of cardiomyocytes and collagen can form an annular cardiomyocyte-collagen strip in the annular groove. After the formed ring-shaped cardiomyocyte-collagen strip was cultured in the ring groove for 7 days, it was taken out and stretched with force, and after 7 days, the samples were taken for morphological and histological detection.
[0010] Results: The constructed cardiomyocyte-collagen strips could beat consistently. Histological studies indicated that the cardiomyocytes in the strips formed a tissue-like tissue re...
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