Canine umbilical cord mesenchymal stem cells

Canine umbilical cord mesenchymal stem cells

|Product attributes:

|Cell Introduction:

Isolation of mesenchymal stem from umbilical cord in dogs; The umbilical cord is a tubular structure in mammals that connects the fetus and placenta. Originally formed by the stalk like elongation of the yolk sac and urinary membrane wrapped around the amniotic membrane. The blood vessels that pass through the urinary membrane in the umbilical cord are the umbilical artery and umbilical vein, while the blood vessels in the yolk sac are the umbilical mesenteric artery and umbilical mesenteric vein. When the yolk sac and its blood vessels degenerate, the umbilical artery and umbilical vein develop, and loose gelatinous stroma can be seen in these gaps. In the uterus, the uterine artery is a capillary that emerges from the maternal part of the placenta and is close to the fetal capillary in the daughter body of the placenta, where it travels between the blood of the mother and the fetusCO2andO2Metabolic products refer to the exchange of metabolic waste and nutrients. The umbilical artery transports waste from the fetus to the placenta, while the umbilical vein transports itO2Nutrients are transported from the placenta to the fetus. Afterwards, metabolic waste from the fetus is transported away by uterine veins, and certain hormones and antibodies are also transferred from the mother to the fetus through the umbilical cord. The umbilical cord contains a large number of stem cells, which are the seeds of life. They differentiate into various cells in the body and produce different fruits - blood cells, nerve cells, bone cells, etc. Stem cells are a population of cells with self-renewal, high proliferation, and multiple differentiation potentials; These cells can maintain their own cellular characteristics and quantity through division, and can further differentiate into various tissue cells, thus playing a positive role in tissue repair and other aspects. 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; Currently, it is mostly used for the treatment of rheumatic and immune diseases. 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.

|Method Introduction:

The canine umbilical cord mesenchymal stem isolated in the laboratory was prepared using the tissue patch method, with a total cell count of approximately5×10⁵cells/Bottle.

|Quality inspection:

Laboratory isolated canine umbilical cord mesenchymal meridianCD90Immunofluorescence 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 canine umbilical cord 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 culture operation:

1) Resuscitate cells:Quickly shake and thaw the cryovial containing 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.

2) Cell passage: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 1ml 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 operating table, 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 a new dish or bottle containing 8ml of culture medium at a ratio of 1:2.

3) Cell cryopreservation:When the cell growth is in good condition, cell cryopreservation can be performed. The following T25 bottles are classified;

When cells are frozen, discard the culture medium, wash with PBS once, and add 1ml. After the cells become round and fall off, add 1ml of serum containing culture medium to terminate digestion, which can be counted using a hemocytometer.

Centrifuge at 1000rpm for 4 minutes to remove the supernatant. Add 1ml of serum to resuspend cells, add serum and DMSO according to the number of cells, gently mix well, DMSO final concentration is 10%, cell density is not less than 1x106/ml, freeze 1ml of cell suspension in each cryovial, pay attention to labeling the cryovial.

3. Place the cryotube in a program cooling box, put it in a -80 degree freezer, and after 2 hours, transfer it to liquid nitrogen storage. Record the location of the freezer for future retrieval.
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|Attention:

1. After receiving non erythroid nucleated cells, first observe whether the cell bottle is intact and whether there is any leakage or turbidity in the culture medium. If any of the above phenomena occur, please contact us in a timely manner.

2. Carefully read the cell instructions to understand cell related information, such as cell morphology, culture medium used, serum ratio, required cytokines, etc., to ensure cell culture conditions. If there are any problems with the cells due to poor cultivation conditions, the responsibility shall be borne by the customer themselves.

3. Wipe the surface of the cell vial with 75% alcohol and observe the cell status under a microscope. Due to transportation issues, a small amount of adherent cells may detach from the bottle wall. Place the cells in a culture box and let them stand for 4-6 hours before removing them for observation. At this point, most cells will adhere to the wall. If the cells still cannot adhere to the wall, please use trypan blue staining to determine cell viability. If it is confirmed that the cell viability is normal, please centrifuge the cells and culture them again with fresh culture medium; If the staining result shows that the cells are inactive, please take a photo and contact us in a timely manner. After confirming the information, we will send it to you again for free.

4. After allowing the cells to adhere to the wall, please pour out the culture medium from the cell bottle and leave 6-8 mL to maintain normal cell culture. When the confluence of ATCC CRL-1711 (Sf9) cells reaches around 80%, normal passage can be achieved.

5. Please ask the customer to use the same culture medium for cell culture under the same conditions. The excess culture medium in the culture bottle can be collected for future use. When cells are passaged, they can be mixed with the customer's own culture medium in a certain proportion to gradually adapt to the culture conditions.

6. It is recommended that customers take several photos of the cells in the first 3 days after receiving them, recording the cell status for communication with the technical department. Due to transportation reasons, some sensitive cells may experience instability. Please contact us promptly to inform us of the specific situation of the cells, so that our technical personnel can track and follow up until the problem is resolved.

7. This cell is for scientific research purposes only