Experimental steps of mouse embryonic stem cells:
1. Remove a tube of cells from liquid nitrogen;
2. Place the cryopreservation tube in a 37 ° C water bath for 2 minutes (or the solution in the tube just completely dissolves)
3. Transfer the cells to a 15ml Falcon tube;
4. Add 5ml ES medium (wash the cryotube with medium);
5. Centrifuge for 3 minutes;
6. Discard the supernatant and resuspend the cells in 2ml ES medium, pipetting at least 10 times;
7. Inoculate in 6-well or 6cm tissue culture dish coated with gelatin (see below);
8. Incubate.
Characteristics of mouse embryonic stem cells:
Alkaline phosphatase staining was positive.
Expression of OCT-4, SSEA-1 and nanog.
The karyotype is normal (40, XY or 40, XX), and the normal karyotype is maintained for a long time.
Teratomas can form in immunodeficient mice.
It has the ability of multi-directional differentiation and can differentiate into cells of three germ layers including neurons.
Quality control of mouse embryonic stem cells:
This product has been tested for bacteria, fungi and mycoplasma.
Mouse embryonic stem cell transportation and preservation:
Use dry ice to save and transport.
When receiving the cells, some of the ice has completely melted, please immediately resuscitate the cells; if there is still dry ice, please immediately put the cells in liquid nitrogen to save for use. High temperature environment.
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