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Human-Primary-Oral-Keratinocytes-Cells

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Human-Primary-Oral-Keratinocytes-Cells

商品詳情參考文獻(xiàn) 相關(guān)資料

	Print Version
Procedure Overview
BioSafety Level	II
Organism	Homo sapiens
Source Organ	Oral Epithelial Tissue (Gingiva)
Growth Properties	Adherent
Morphology	Polygonal
Image
Cell Type Characterization
Passage Number	2
Population Doubling	10
Applications	For Research Use Only
Disease	Normal
Propagation	These cells grow well on Poly-L-Lysine coated flasks. A Poly-L-Lysine Coating Solution (0.1%) is available at abm (TM061). The base medium for this cell line is Prigrow X series medium available in ABM, Cat. No. TM4074. Atmosphere: air, 95%; Carbon dioxide (CO2), 5%.
Subculturing	1. Subculture when the culture reaches 90% confluency or above. 2. Prepare Poly-L-Lysine coated culture flask (2 μg/cm square) one day before subculture. 3. Warm complete medium, TrypsinEDTA solution, TNS, and DPBS (Ca++ and Mg++ free) to room temperature. We do not recommend warming reagents and medium in a 37°C water bath prior to use. 4. Rinse the cells with DPBS. 5. Add 8 ml of DPBS and then 2 ml of TrypsinEDTA solution into flask. Gently rock the flask to ensure complete coverage of cells by T/E solution. Incubate the flask in a 37°C incubator for 1 to 2 minutes or until cells completely round up. Use a microscope to monitor the change in cell morphology. 6. During incubation, prepare a 50 ml conical centrifuge tube with 5 ml of FBS. 7. Transfer T/E solution from the flask to the 50 ml centrifuge tube (a small percent of cells may detach) and continue to incubate the flask at 37°C for another 1 to 2 minutes (no solution in the flask at this moment). 8. At the end of incubation, gently tap the side of the flask to dislodge cells from the surface. Check under a microscope to make sure that all cells detach. 9. Add 5 ml of TNS to the flask and transfer detached cells to the 50 ml centrifuge tube. Rinse the flask with another 5 ml of TNS to collect the residual cells. 10. Examine the flask under a microscope for a successful cell harvest by looking at the number of cells being left behind? there should be less than 5%. 11. Centrifuge the 50 ml centrifuge tube at 1000 rpm for 5 minutes. Resuspend cells in culture medium. 12. Count and plate cells in a new Poly-L-Lysine coated culture vessel.
Preservation	1. Freeze Medium: Complete growth medium with 20% FBS and 10% DMSO. 2. Storage Temperature: Liquid nitrogen vapour phase.
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.