Duck embryonic liver mesenchymal stem cells

Cell Introduction:

Duck embryonic liver mesenchymal stem isolated from embryonic liver tissue; The liver is an organ in the body primarily responsible for metabolic functions, and plays a role in the synthesis of antioxidant, glycogen storage, and secreted proteins; The liver also produces bile in the digestive system. The liver is a large organ in the internal organs of the body, located in the abdomen below the right diaphragm, at the anterior end of the gallbladder and in front of the right kidney, above the stomach. The liver is a large digestive gland in the body's digestive system, which is reddish brown in colorVCharacter shaped organ. The liver is the main organ for urea synthesis and an important organ for metabolism. mesenchymal stem cellMSC）It is a type of stem cell with high self-renewal and multi-directional differentiation potential. Under different induction conditions, it can differentiate into various tissue cells other than hematopoietic cells, and has functions such as hematopoietic support, immune regulation, and tissue repair. mesenchymal stem cellMSCs）It is an adult stem cell with self-renewal and multipotent differentiation potential, which exists in bone marrow, adipose tissue, umbilical cord blood, and various fetal tissues. It can secrete various cytokines and growth factors, promoting hematopoietic stem cells（HSC）Proliferation and differentiation.MSCsIt also has immunomodulatory, anti-inflammatory, and tissue repairing effects, which can alleviate graft-versus-host disease（GVHD）And other transplant related complications. Non parenchymal cells in embryonic liver containMSCAnd with a large quantity, it can become seed cells.

Method Introduction:

Laboratory isolated duck embryonic liver mesenchymal stem was used-Prepared by collagenase combined digestion method, with a total cell count of approximately5×10⁵cells/Bottle.

Quality inspection:

Laboratory isolated duck embryonic liver mesenchymal stem cellsCD44Immunofluorescence identification, purity can reach90%Above, and not containingHIV-1TheHBVTheHCVMycoplasma, bacteria, yeast, fungi, etc.

Training Information:

Culture medium: containingFBSTheEGFThebFGFThePenicillinTheStreptomycinwait

Fluid change frequency: every2-3Change the fluid once every day

Growth characteristics: Wall adhesion

Cell morphology: fibroblast like

Passage characteristics: passable3-5Dai Zuo

Digestive fluid:0.25%

Cultivation conditions: Gas phase: Air,95%；CO2，5%

The in vitro culture period of duck embryonic liver mesenchymal stem is limited; It is recommended to use specialized growth medium and correct operating methods to cultivate the cells, in order to ensure the optimal cultivation state of the cells.

Cell passage steps:

If the cell density reaches 80% -90%, subculture can be carried out.
1. Discard the culture supernatant and rinse the cells 1-2 times with PBS that does not contain calcium or magnesium ions.
2. Add 2 ml of digestion solution (0.25% Trypsin 0.53mM EDTA) to a culture bottle and place it in a 37 ℃ incubator for digestion for 1-2 minutes. Then observe the cell digestion under a microscope. If most of the cells become round and fall off, quickly take them back to the workstation, tap the culture bottle a few times, and add a small amount of culture medium to terminate digestion.
3. Add 6-8ml/bottle of culture medium, gently mix and aspirate, centrifuge at 1000RPM for 4 minutes, discard the supernatant, add 1-2mL of culture medium and blow evenly.
4. Divide the cell suspension into new dishes or bottles containing 8ml of culture medium in a ratio of 1:2 to 1:5.

Cell recovery steps:

Quickly shake and thaw the cryovial containing 1mL of cell suspension in a 37 ℃ water bath, then add 4mL of culture medium and mix well. Centrifuge at 1000RPM for 4 minutes, discard the supernatant, add 1-2mL of culture medium, and blow evenly. Then add all cell suspensions to the culture bottle and culture overnight (or add the cell suspensions to a 10cm dish, add about 8ml of culture medium, and culture overnight). The next day, change the solution and check the cell density.

Cell cryopreservation steps:

When the cell growth is in good condition, cell cryopreservation can be performed. Taking T25 bottles as an example below;
When cells are frozen, discard the culture medium, wash the bottom of the bottle 1-2 times with PBS, and then add 1ml. After the cells become round and fall off, add 2ml of culture medium to terminate digestion, which can be counted using a hemocytometer.
Centrifuge at 2000 RPM for 5 minutes to remove the supernatant. Resuspend with serum and add DMSO to a final concentration of 10%. After adding DMSO, quickly mix well and distribute it into cryovials in quantities of 1ml each. Pay attention to labeling the cryovials properly. Our company freezes more than 1X106 cells per freezing tube.
3. Place the cryovial in a program cooling box and store it in a -80 degree freezer for at least 2 hours before transferring it to liquid nitrogen storage. Record the location of the freezer for future retrieval.
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