Human Primary Pancreatic Islets
| Cat. No. | T0199 |
| Name | Human Primary Pancreatic Islets |
| Description |
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. Cells are shipped as live cells and cannot be cryopreserved. Experiments should be conducted within 7 days upon cell arrival. These cells are an ideal model for islet grafting survival studies and can provide insight into development of novel therapies for other pancreas related diseases. |
| Organism | Human (H. sapiens) |
| Tissue | Pancreas |
| Donor History | Normal tissue |
| Growth Properties | Suspension, round |
| Storage Condition | Vapor phase of liquid nitrogen, or below -130°C. |
| Shipping Conditions | Ship with dry ice. |
| Product Format | Frozen |
| Intended 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. |
| BioSafety | II |
| Certificate of Analysis | For batch-specific test results, refer to the applicable certificate of analysis that can be found at www.abmgood.com. |
| Growth Conditions |
Refer to detailed instructions outlined below for islet handling. The media supplied during shipment is not sufficient for islet maintenance. TM0199 (ready-to-use) is recommended for islet maintenance. Note: 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. Instructions for Islet Recovery After Shipment Receipt 1. Cool the TM0199 (supplied as a complete media) at 6-10°C refridgerator prior to receiving the islets. 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. 3. Break up clumps by gently pipetting the islets and ensure islets are well dispersed. Completely cover the islets with TM0199. Centrifuge 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. 6. Check viability and perform a cell count. 5. Culture cells at 10K IEQ/T150 flask with 40 mL of complete TM0199 media. 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. 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. 5. Incubate cells according to recommended conditions. Instructions for Short Term Islet Culture (37°C 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. Instructions for Long Term Islet Storage (8°C refrigerator/incubator) 1. Seal caps of islet culture vessels with parafilm and place in 8°C refrigerator/incubator. 2. Media must be changed every 7th day following the instructions for islet media change (above). However, all media used should be cold (cool it in a 2-8°C refrigerator prior to use). 3. Pleace the flasks in a 37°C incubator, with 5% CO2 overnight. 4. Repeat step 1. 5. Islets can be cultured for up to a month using this method. |
| Unpacking and Storage Instructions |
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°C until ready to be washed. To ensure the highest level of viability initiate culture as soon as possible upon receipt. |
| Subculture Protocol |
abm does not recommend subculturing this cell line. We advise using cells for experimental purposes as soon as possible after intiating culture. |
| Warranty | 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”. |
| 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 licensing@abmgood.com. 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 μg, Cat.# C207, $450.00) or cell lysate (100 μg, 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." |
| Application | Research Use Only. |
| Material Citation | If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. T0199 |
- Important Considerations for Primary Cells
- Primary Cell Handling Instructions Upon Arrival
- Protocol for Islet Culturing
- Protocol for Islet Media Change
- Protocol for Recovering Islets after receiving in Transport Solution
- Important Considerations for Cell Culture
- Cell Handling Instructions Upon Arrival
| I want to make sure these cells express my gene of interest before I decide to buy the cell line. Can you provide a sample so this can be tested? | |
|
We do not perform extensive downstream characterization, or gene expression profiling of our cell lines. However, you can check the ‘References’ tab to see if a publication is available, or you can contact us to see if RNA or cell lysates are available for your cell line of interest. Please directly contact us for further information at quotes@abmgood.com.
|
| Does abm's PriGrow series medium contain antibiotics? | |
|
No, the medium does not contain antibiotics and needs to be supplemented by the end-user as desired.
|
| Do I need to use abm's media and Applied Cell Extracellular Matrix (G422) to culture my cells? | |
|
We strongly encourage using the recommended media and culture conditions described in the "Growth Conditions" section as they have been optimized for cell growth. Other supplier's media and coating conditions have not been tested.
|
| What is the difference between Applied Cell Extracellular Matrix (G422) and Collagen Coating? | |
|
The main component of our Applied Cell Extracellular Matrix (G422) is type I collagen specifically. For more information on abm's Applied Cell Extracellular Matrix please visit: http://www.abmgood.com/Applied-Cell-Extracellular-Matrix-G422.html.
|
| Do I have to use T25 ECM-coated flasks for growing the cells? | |
|
We strongly recommend thawing cells in the T25 flasks specified on the cell-specific product page to ensure optimal post-thaw recovery of the frozen cells before testing other culture vessels.
|
| If I want to plate these cells to multi-well plates (e.g. 96 well plates) or dishes, how should I prepare the plates? | |
| What percentage of Trypsin-EDTA should be used to subculture cells? Can I use Trypsin-EDTA containing phenol red? | |
|
The standard concentration used for subculturing is 0.25% Trypsin-EDTA, available at abm (Cat. No. TM050). Yes, Trypsin-EDTA containing phenol red can be used as it does not interfere with trypsinization.
|
| How often do I need to change the media? | |
|
Media should be changed every 2-3 days or as specified within the recommended growth conditions.
|
| Why are these cells classified as biosafety level II? | |
|
We follow the CDC-NIH recommendations that all mammalian sourced products should be handled at the Biological Safety Level 2 to minimize exposure of potentially infectious products. This information can be found in 'Biosafety in Microbiological and Biomedical Laboratories' (1999). Your institution's Safety Officer or Technical Services will be able to make the call as to whether BioSafety Level I is possible with these cells at your site, if desired.
|
| How long can I store frozen vials? | |
|
Cells that are properly frozen using an effective cryoprotective agent can be stored in liquid nitrogen vapor phase (or below -130°C) indefinitely without affecting viability.
|
| What is the recommended storage temperature for cells? | |
|
In general, if you received:
|
| Can I substitute heat-inactivated FBS with FBS or vice versa? | |
|
FBS and heat-inactivated FBS are different in their composition; they cannot be substituted for one another.
|
| My cells are not detaching, what method do you recommend to trypsinize the cells? | |
|
| How are live cells shipped? | |
|
Live cells are shipped as a live culture in a T12.5 flask. Cells are seeded at an optimal confluency to avoid metabolic waste build up during transport. In addition, we thoroughly test our cell lines for functional freeze-thaw recovery. For more information on how to handle live cells, please view our detailed instructions here (https://www.abmgood.com/uploads/document/Cell_Handling_Instructions_Upon_Arrival_150623.pdf).
|
| Why are cells not attaching and forming clumps after subculturing with trypsin? | |
|
Cells form clumps and display attachment issues when the trypsinization agent is not effectively neutralized. To completely neutralize the trypsin agent, add an equal amount of complete growth media prior to centrifugation. Subtle changes in culture conditions, particularly in pH and the quality of serum used in the growth medium, may also affect the amount of clumping exhibited by the cells.
|
| Why are my cells forming clumps after plating? | |
|
It is important to ensure cell clumps are broken up through gentle re-suspension by pipetting up and down several times after pelleting cells by centrifugation. This will ensure a homogenous single cell suspension for even plating.
|
| Do I need to add the selection drug to the complete growth medium? | |
|
We recommend maintaining selection pressure using the drug concentration specified in the Growth Conditions section.
|
| Where can I find the CoA for my product? | |
|
Certificates of analysis can be found here (https://www.abmgood.com/CoA-Library.html).
|
|
What is your warranty or return policy? |
|
|
Our warranty and return policy is outlined in abm’s Terms and Conditions, including details on product quality, limitations, and claims. For additional questions, our Order team is happy to assist and can be reached at order@abmgood.com. |
|
How many times can cells divide? |
|
|
The number of times cells can divide depends on the cell type:
|
|
Do I need Applied Cell Extracellular Matrix (G422) if I am using PriCoat™ flasks? |
|
|
Please refer to the Growth Conditions section of your specific product page. This section will indicate whether Applied Cell Extracellular Matrix (G422), PriCoat™ flasks, or both are recommended for optimal cell attachment and growth, as requirements may vary by cell type. |
- Verma, G et al. "JNK1/2 regulates ER-mitochondrial Ca2+ cross-talk during IL-1β-mediated cell death in RINm5F and human primary β-cells" Mol Biol Cell 24(12):2058-71 (2013). DOI: 10.1091/mbc.E12-12-0885. PubMed: 23615449.
- Han, Z. et al. "Survivin silencing and TRAIL expression using oncolytic adenovirus increase anti-tumorigenic activity in gemcitabine-resistant pancreatic cancer cells" Apoptosis :1-14 (2015). PubMed: 26677013.
- Na, Y et al. "Potent antitumor effect of neurotensin receptor-targeted oncolytic adenovirus co-expressing decorin and Wnt antagonist in an orthotopic pancreatic tumor model" Journal of Controlled Release Part B:766–782 (2015). DOI: doi:10.1016/j.jconrel.2015.10.015. Application: cell-based assay.
- Lee, JM et al. "A new synthetic 2′-hydroxy-2,4,6-trimethoxy-5′,6′-naphthochalcone induces G2/M cell cycle arrest and apoptosis by disrupting the microtubular network of human colon cancer cells" Cancer Lett 354(2):348-54 (2014). DOI: 10.1016/j.canlet.2014.08.041. PubMed: 25193463.
- Na, Y et al. "Potent antitumor effect of neurotensin receptor-targeted oncolytic adenovirus co-expressing decorin and Wnt antagonist in an orthotopic pancreatic tumor model" J. Controlled Release 220:766-782 (2015). DOI: http://dx.doi.org/10.1016/j.jconrel.2015.10.015. Application: Virus effects.