Adipose-derived stem cells-ecm modified sis composite engineered bone and its preparation method
An adipose stem cell and human adipose stem cell technology, which is used in medical science, prosthesis, tissue regeneration, etc., can solve problems such as uncontrollable factors, and achieve the effects of good safety, high mechanical strength, and excellent biocompatibility.
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Embodiment 1
[0027] The preparation method of the fat stem cell-ECM modified SIS composite engineered bone of embodiment 1 comprises the following aseptic operation steps:
[0028] 1) Osteoblast culture and passage: Prepare α-MEM medium containing 10% fetal bovine serum as the osteoblast culture medium; place MC3T3-E1Subclone 14 cells isolated from the MC3T3-E1 cell line of the anterior cranial bone of the mouse into Cultivate in a cell culture incubator with 5% carbon dioxide concentration and 37°C, discard the old culture medium every 48 hours and add an equal amount of new culture medium, and carry out cell passage when the confluence of MC3T3-E1Subclone 14 cells is greater than 90% , the process of subculture of the cells is as follows: discard the old culture medium, add PBS buffer solution without calcium and magnesium ions, and wash twice; then add 1 mL of 0.25 wt% trypsin containing EDTA, and incubate at 37°C for 1 min; Collect the cells and add them to a centrifuge tube, centrifug...
Embodiment 2
[0032] The preparation method of the fat stem cell-ECM modified SIS composite engineered bone of embodiment 2 comprises the following aseptic operation steps:
[0033] 1) Osteoblast culture and passage: Prepare α-MEM medium containing 10% fetal bovine serum as the osteoblast culture medium; place MC3T3-E1Subclone 14 cells isolated from the MC3T3-E1 cell line of the anterior cranial bone of the mouse into Cultivate in a cell culture incubator with 5% carbon dioxide concentration and 37°C, discard the old culture medium every 48 hours and add an equal amount of new culture medium, and carry out cell passage when the confluence of MC3T3-E1Subclone 14 cells is greater than 90% , the process of subculture of the cells is as follows: discard the old culture medium, add PBS buffer without calcium and magnesium ions, wash 3 times; then add 1.5mL of 0.25wt% trypsin containing EDTA, and incubate at 37°C for 1min ;Collect the cells and add them to the centrifuge tube, and centrifuge at 1...
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