Primer and method for detecting Nanog gene expression quantity during mesenchymal stem cell passage
A technology of gene expression and qualitative stem cells, which is applied in the detection field of primers and detection of Nanog gene expression in the process of passage of mesenchymal stem cells, can solve the problems of low sensitivity, low detection efficiency, poor accuracy, etc., and achieve high sensitivity High, good primer specificity, good accuracy
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[0025] A method for detecting the amount of Nanog gene expression during in vitro passage of mesenchymal stem cells, comprising the following steps:
[0026] (1) Extraction of mesenchymal stem cells
[0027] UC-MSCs: After rinsing the umbilical cord with normal saline in a biological safety cabinet, peel off the amniotic epithelium, remove the umbilical artery and umbilical vein, then cut the umbilical cord jelly into pieces and add 30mL type Ⅱ collagenase, place at 37°C, 200r The digestion was continued for 6 h at 100 mesh sieve, and then centrifuged at 1200 r / min for 10 min, the supernatant was discarded, and the cells were gently washed with D-Hank's solution for 3 times, and the cells were resuspended in serum-free medium (hyclone company). 5% CO at 37°C 2 After culturing in a constant temperature incubator for 6-7 days, change the medium completely, discard non-adherent cells, and separate UC-MSCs from other tissue cells according to the difference in adhesion. After sep...
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