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Enhanced Primary Human Liver Sinusoidal Endothelial Cells

SKU : BHC10900002

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Applied Biological Materials (abm) Inc.

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Mfr.No.	T5997
Description	Enhanced Primary Human Liver Sinusoidal Endothelial Cells are expanded primary cells that retain the properties of primary liver sinusoidal endothelial cells allowing for a reliable in vitro?esearch model. These cells are ready-to-use for applications in research and development, such as viral infections, screening, co-cultures, transient transfection, and for toxicity studies. The cells offer a unique ability to be kept in longer term culture when compared to standard primary liver sinusoidal endothelial cells which does not proliferate well,?n vitro. The cells are ready-to-use after thawing. abm recommends passaging cells 1-2 times; beyond this limit cells may exhibit phenotypic changes and express senescence genes.
Cell Type	Primary Cells
Biosafety Level	2
Disclaimer	1. Sale of this item is subjected to the completion of a Material Transfer Agreement (MTA) by the purchasing individual/institution for each order. If you have any questions regarding this, please contact us at [email protected]. 2. 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 or ordering RNA (0.5 ?g, Cat.# C207, $450.00) or cell lysate (100 ?g, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final. 3. We 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). 4. 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. 5. 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. 6. abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the "Warranty Period."
Expression Region	MMR+, CD31+
Gender	Donor Info Not Disclosed
Growth Properties	Adherent, endothelial-like
Growth Conditions	PriCoat?ECM T25 Flasks (G999) or Applied Cell Extracellular Matrix (G422) are?equired?or cell adhesion to the culture vessels.?uman Liver Sinusoidal Endothelial Cell Thawing Medium (TM111) + Human Liver Sinusoidal Endothelial Cell Media Kit (TM112), 37.0?, 5% CO?.Note: TM111 is required for optimal recovery of cells post-thaw. Cells are grown using TM112.
Quantity	5x105 cells / 1.0 ml
Tissue	Liver
Morphology	Endothelial-like
Notes	1. Visually examine the packaging containers for signs of leakage or breakage. 2. Immediately transfer frozen cells from dry ice packaging to a temperature below -130?, preferably in liquid nitrogen vapor phase storage, until ready for use. To ensure the highest level of viability, thaw the vial and initiate culture as soon as possible upon receipt. If continued storage is desired, the vial should only be stored below -130? or in liquid nitrogen vapor phase. Do not store at -70?, as it will result in loss of viability.
Organism	Human (H. sapiens)
Species	Human (H. sapiens)
Propagation	Use of PriCoatTM 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 unless otherwise specified in the Propagation Requirements below.Use the ready-to-use Human Liver Sinusoidal Endothelial Cell Thawing Medium (TM111) and the Human Liver Sinusoidal Endothelial Cell Media Kit (TM112) which comes with the Human Liver Sinusoidal Endothelial Cell Basal Medium, and Human Liver Sinusoidal Endothelial Cell Supplement Mix available from abm.Carbon dioxide (CO2): 5%, Temperature: 37.0°C.To make complete Human Liver Sinusoidal Endothelial Cell Growth Medium add the entire content of Human Liver Sinusoidal Endothelial Cell Supplement Mix into Human Liver Sinusoidal Endothelial Cell Basal Medium and mix properly in BioSafety Cabinet. Addition of the Human Liver Sinusoidal Endothelial Cell Supplement Mix may make medium appear more opaque.To Thaw:1. Pre-warm Human Liver Sinusoidal Endothelial Cell Thawing Medium and fully supplemented Human Liver Sinusoidal Endothelial Cell Growth Medium to room temperature.2. Carefully remove cryovial from storage tank.3. Thaw cells in 37°C water bath until only a small chunk of ice is left. Do not shake the vial, or take it out of the water during thawing, as this will damage the cells.4. Spray the vial and the tube containing 50 ml of thawing medium with 70% ethanol and transfer to a Biosafety cabinet.5. Transfer the now completely thawed cell suspension from the cryovial into 10 ml Human Liver Sinusoidal Endothelial Cell Thawing Medium in a new 50 ml conical tube by gently pipetting the cells into the medium using a 2 ml pipette.6. Use a 1 ml pipette to transfer 1 ml of the thawing medium with the cells back to the cryovial and pipette the contents back into the 50 ml tube. Repeat this process twice to completely remove the cells from the cryovial.7. Pellet the cells by centrifuging at 620×g for 5 min at RT. Important note: Higher g-forces will significantly reduce cell recovery.8. Aspirate the supernatant without disturbing the pellet. Leave approximately 200 mul medium on top of the cells.9. Add 800 mul of the fully supplemented Human Liver Sinusoidal Endothelial Cell Growth Medium and gently loosen and re-suspend the cells by pipetting them up and down 1-2 times. Do not vortex or shake the cells as this will compromise cell survival.11. Determine viable cell number by cell count.12. Dilute the Enhanced Primary Human Liver Sinusoidal Endothelial Cells in pre-warmed, fully supplemented Human Liver Sinusoidal Endothelial Cell Growth Medium and seed at ~10,000 cells/cm2 in ECM-coated cell culture flasks (G422) or appropriate cell culture dishes.13. Incubate the cells at 95% humidity, 37°C and 5% CO2.14. Next day, add fresh new Human Liver Sinusoidal Endothelial Cell Growth Medium. Conduct complete media change every other day with fresh Human Liver Sinusoidal Endothelial Cell Growth Medium for optimal growth.
Cryopreservation	Cryopreservation Medium (TM024), or complete growth media with 10% DMSO.

Applications Range	Research Use Only.
Applications	For Research Use Only

Shipping	Ship with dry ice.
Product Format	Frozen
Product Use	This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
Storage	Vapor phase of liquid nitrogen, or below -130?.

Thawing Protocol	Pre-warm Human Liver Sinusoidal Endothelial Cell Thawing Medium (TM111) and fully supplemented Human Liver Sinusoidal Endothelial Cell Growth Medium (TM112) to room temperature. 1.?haw cells quickly in a 37? water bath while agitating gently (maximum 2 minutes). The vial cap should be kept above the water level to minimize the risk of contamination. Do not shake vial. 2. Decontaminate the vial by spraying and wiping the exterior of the vial with 70% ethanol. From this point onwards, all operations should be strictly carried out inside a biological safety cabinet using aseptic conditions. 3. Transfer the cell suspension into a 50ml sterile conical tube containing 20ml of pre-warmed, Human Liver Sinusoidal Endothelial Cell Thawing Medium (TM111). Centrifuge cells at 280? for 5 minutes.? 4. Aspirate the supernatant without disturbing the cell pellet. Re-suspend the cell pellet in 1 ml of pre-warmed, complete Human Liver Sinusoidal Endothelial Cell Growth Medium (TM112) and seed at?0,000 cells/cm2?n the appropriate G422 coated culture vessel. 5. Incubate the cells at the recommended conditions. 6. After 24 hours, add fresh Human Liver Sinusoidal Endothelial Cell Growth Medium (TM112).?Change media?very other day with fresh Human Liver Sinusoidal Endothelial Cell Growth Medium (TM112) until cells become 70-80% confluent.
Subculture Protocol	Cells are sensitive to trypsin; TrypLE Express is recommended for subculture procedures.?Cells can be passaged for one subculture once they become 70-80% confluent. Do not expand cells furthers.?Volumes given below are for a T75 flask; proportionally increase or decrease the volume as required per culture vessel size. 1. Aspirate the culture media, wash the adherent monolayer with 1X PBS, then and add 2-3ml of pre-warmed TrypLE Express to the culture vessel. 2. Observe the cells under a microscope to confirm detachment (typically within 2-10 minutes). Cells that are difficult to detach can be put in 37?, for several minutes to facilitate detachment. 3. Neutralize TrypLE Express by adding an equal volume of the complete growth media into the culture vessel. 4. Transfer the culture suspension into a sterile centrifuge tube, and centrifuge at 280xg for 5 minutes. The actual centrifuge duration and speed may vary depending on the cell type. 5. Aspirate the supernatant, and re-suspend the pellet with pre-warmed fresh complete growth media. Seed cells at 10,000 cells/cm2?n the appropriate G422 coated culture vessel. 6. Incubate the cells at the recommended conditions. 7. Allow the cells to expand over the next 2-3 days then perform experimental assays as desired.

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SKU	Supplier No.	Size	Your price	Quantity
BHC10900002	T5997	5x10^5 cells / 1.0 ml	-		Add to Quote

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