Fish spleen lymphocytes

Fish spleen lymphocytes

|Product attributes:

|Cell Introduction:

Separation of fish spleen lymph from spleen tissue; The spleen is a large immune organ in the body, accounting for the total amount of lymphoid tissue in the body25%It contains a large number of lymphocytes and macrophages, and is the center of cellular and humoral immunity in the body. Located deep in the outer rib arch behind the left rib area, and9－11Rib relative, long axis and th10The ribs are consistent. The diaphragm is adjacent to the diaphragm and left costophrenic sinus, with the stomach in front and the left kidney and adrenal gland in the back. The lower end is adjacent to the splenic groove of the colon, and it is a soft reticular endothelial cell organ. Lymphocytes（lymphocyte）A type of white blood cell produced by lymphoid organs and an important cellular component of the body's immune response function. According to their differences in occurrence and function, lymphoid organs can be divided into two categories: central lymphoid organs (also known as primary lymphoid organs) and peripheral lymphoid organs (also known as secondary lymphoid organs). The former includes the thymus, supraluminal sac, or their equivalent organs (some believe it is the bone marrow in mammals). They can continuously proliferate lymphocytes without antigen stimulation and transfer them to surrounding lymphoid organs after maturation. The latter includes the spleen, lymph nodes, etc. Mature lymphocytes rely on antigen stimulation to differentiate and proliferate, thereby exerting their immune function.

|Method Introduction:

The fish spleen lymph isolated in the laboratory was prepared using a combination of mechanical grinding and density gradient centrifugation, with a total cell count of approximately5×10⁵cells/Bottle.

|Quality inspection:

The fish spleen lymph isolated in the laboratory has been tested and does not containHIV-1TheHBVTheHCVMycoplasma, bacteria, yeast, fungi, etc.

|Training Information:

Culture medium: containingFBSThePenicillinTheStreptomycinwait

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

Growth characteristics: suspended

Cell morphology: Round

Passage characteristics: non proliferative; Not passaged

Digestive fluid:0.25%

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

The in vitro culture period of fish spleen lymph 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.

|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.

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