Human Primary Pancreatic Islets can be split into different culture vessels (i.e. multi-well plates) for experimental needs, but they will not proliferate in vitro. Experiments should be conducted within 7 days upon cell arrival. Upon thaw, the cells cannot be cryopreserved.These cells are an ideal model for long-term islet grafting survival studies and can provide insight into development of novel therapies for other pancreas related diseases.
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."
Donor History
Normal tissue
Gender
Donor Info Not Disclosed
Growth Properties
Suspension, round
Growth Conditions
Refer to detailed instructions in the following sections for islet handling:?npacking and Storage Instructions, Thawing Protocol, Subculturing Protocol,?ryopreservation.?he media supplied during shipment is not sufficient for islet maintenance. TM0199 (ready-to-use) is recommended for islet maintenance.?ote: Human Primary Pancreatic Islets can be split into different culture vessels (i.e. multi-well plates) for experimental needs, but they will not proliferate in vitro. Experiments should be conducted within 7 days upon cell arrival. Cells cannot be cryopreserved post-thaw.
Quantity
5x105 cells / 1.0 ml
Tissue
Pancreas
Morphology
Cobble-stone
Notes
1. Visually examine the packaging containers for signs of leakage or breakage. 2. Islet recovery from transport requires Islet Recovery Media (Cat. TM0199-A). Freshly received islets can be stored at 6-10? until ready to be washed.?To ensure the highest level of viability, 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 PriCoatTMT25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells inECM-coated culture vessels unless otherwise specified in the Propagation Requirements below.The base medium for this cell line is Prigrow X series medium available at abm, Cat. No. TM104. Cells must be grown on gelatin coated plates. Gelatin Coating Solution (0.1%) is available at abm, TM063. Cells can only propagate for a maximum of 2 passages at split ratio of 1:2. Recommend to conduct media change every 3rd day to maintain optimal growth. Carbon dioxide (CO2): 5%, Temperature: 37.0
Cryopreservation
Instructions for Long Term Islet Storage (8? refrigerator/incubator) 1. Seal caps of islet culture vessels with parafilm and place in?? refrigerator/incubator.?. Media must be changed every 7th day following the instructions for islet media change (above). However, all media used should be?old?cool it in a 2-8??efrigerator prior to use).3. Pleace the flasks in a 37? incubator, with 5% CO2?vernight.4. Repeat step 1.?. Islets can be cultured for up to a month using this method.
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
Instructions for Islet Recovery After Shipment Receipt 1. Cool the TM0199 (supplied as a complete media) at 6-10? refridgerator prior to receiving the islets.?br />2. Pre-wet centrifuge tubes with TM0199 media and transfer islets from the transport bottle to the centrifuge tubes. Ensure all islets are remove by washing the transport bottle with media at least 2 times.?br />3. Break up clumps by gently pipetting the islets and ensure islets are well dispersed. Completely cover the islets with TM0199.?entrifuge cells at 180xg for 2 minutes. 4. Aspirate the supernatant, leaving behind 2-3 ml. Disrupt the pellet by gently tapping the sides of the centrifuge tube. Add 5 mL of fresh TM0199 and break up any visible clumps through gente pipetting. 5. Add additional TM0199 to the desired amount required for the appropraite culture vessel.?br />6. Check viability and perform a cell count. 5. Culture cells at 10K IEQ/T150 flask with 40 mL of complete TM0199 media.
Subculture Protocol
Instructions for Islet Media Change 1. Warm TM0199 media to room temperature.? 2. Angle culture vessel for at least 30 minutes to allow islets to congregate on one corner of the culture vessel. Test media after 30 minutes to ensure islets more than 50 microns in size are not visible. Allow for additional settling time until no islets are present in test sample.?br>3. Maintain flask position and aspirate 50% of the used media without disturbing the congregated islets. 4. Add the same volume of fresh TM0199 that was removed.?br>5. Incubate cells according to recommended conditions.?Instructions for Short Term Islet Culture (37? incubator, 5% CO2) 1. For cells in short-term culture, media should be changed every 2-3 days.? 2. Assess viability and purity each time media is changed. Islets can be cultured up to 2 weeks using this method.