Human-Normal-Dendritic-Cells
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Human-Normal-Dendritic-Cells
Human-Normal-Dendritic-Cells
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Print Version |
| Description |
Human Dendritic cells (DCs) are immune cells that form part of the mammalian immune system. Their main function is to process antigen material and present it on the surface to other cells of the immune system, thus functioning as antigen-presenting cells. They act as messengers between the innate and adaptive immune systems. Dendritic cells are present in small quantities in tissues that are in contact with the external environment, mainly the skin and the inner lining of the nose, lungs, stomach and intestines. They can also be found in an immature state in the blood. Once activated, they migrate to the lymphoid node where they interact with T cells and B cells to initiate and shape the adaptive immune response. |
| Procedure Overview |
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| BioSafety Level |
II |
| Organism |
Homo sapiens |
| Source Organ |
Blood |
| Isolation |
These cells are extracted from whole blood using Lymphoprep(R), a hydrophilic polysaccharide that separates layers of blood, with monocytes and lymphocytes forming a buffy coat under a layer of plasma. |
| Growth Properties |
Adherent |
| Morphology |
Multipolar |
| Passage Number |
They are cryopreserved immediately after isolation |
| Population Doubling |
Dendritic cells do not proliferate in vitro |
| Recommended Seeding Density |
40,000 cells /cm2 |
| Markers |
CD123, BDCA-2, BDCA-4 |
| Applications |
For Research Use Only.The most common applications of dendritic cells are dendritic cell – T cell activation assays. |
| Disease |
Normal |
| Propagation |
Use of PriCoat? T25 Flasks (G299) or . Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells in ECM-coated culture vessels with the following conditions. The medium for this cell line is Prigrow X series medium available in abm, Cat. No. TM4021.
Atmosphere: air, 95%; Carbon dioxide (CO2), 5%. Temperature: 37.0°C.
Thawing Human Primary Dendritic Cells:
1. Prior to thawing cells, prepare a thawing medium by adding 10 ml of fetal bovine serum (TM999) to 40 ml of TM4021 basal medium OR by adding 1 ml of bovine serum albumin (BSA) to 49 ml of TM4021 basal medium. Allow thawing medium to equilibrate to room temperature.
2. Quickly thaw the frozen vial in a 37°C water bath being careful not to submerge the entire vial. When the last sliver of ice melts, remove it. Do not submerge the vial completely. Thawing the cells for longer than 2 minutes results in less than optimal results.
3. Using a micropipette, gently add the thawed cell suspension to a 50 ml sterile polypropylene
centrifuge tube. Rinse the vial with 1.0 ml of thawing medium and add the rinse to the cell
suspension drop by drop while gently swirling the tube. This step should take approximately 1
minute.
4. Slowly bring the total volume of cell suspension up to approximately 35 ml by adding room
temperature thawing medium slowly, while gently swirling the tube.
5. Centrifuge at 1000 rpm for 5-8 minutes at room temperature.
6. Carefully remove all but approximately 1ml of the supernatant by pipette. Resuspend the pellet in the remaining medium using a micropipette. Avoid excessive pipetting.
7. Transfer the cell suspension to a new 15 ml sterile polypropylene centrifuge tube. Rinse the
previously used 50 ml polypropylene centrifuge tube with approximately 5 ml of thawing medium and add the rinse to the cell suspension slowly while gently swirling the tube.
8. Slowly bring the total volume of cell suspension up to approximately 14 ml by adding room
temperature thawing medium slowly while gently swirling the tube.
9. Centrifuge at 1000 rpm for 5-8 minutes at room temperature.
11. Carefully remove all but approximately 200 μl of the supernatant by pipette.
12. Dilute the cells to a final volume of 1 to 2 ml using complete medium TM4021 and note the total volume of the diluted cell suspension. Resuspend the pellet in the medium using a micropipette. Avoid excessive pipetting.
13. Count and plate cells into the designated culture vessel.
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| Quality Control |
abm’s cryopreserved PBMCs consistently exhibited viability above 87% (Day 0 when thawed) with less than a 7.5% deviation from that of fresh PBMCs. |
| Disclaimer |
1. The CoA for this product (provided upon request) verifies the cell-type specific gene expression via RT-PCR only. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers. All sales are final.
2. We strongly recommend live cell shipments for ease of cell transfer and this option can be requested at the time of ordering. Please note that the end-user will need to evaluate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical location. In addition, we thoroughly test our cell lines for freeze-thaw recovery. If frozen cells were received and not recovered in your lab under the exact, specified conditions (using recommended culture vessel, media, additional supplements, and atmospheric conditions), a live cell replacement is possible at a cost (plus shipping).
3. All of abm's cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic application(s). Please contact a technical service representative for more information.
4. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature and provided for informational purposes only. abm does not warrant that such information has been shown to be accurate. |
