Jun 11, 2020
Version 2
Packaging and Cold Shipping of Human Islets V.2
- Integrated Islet Distribution Program1
- 1Integrated Islet Distribution Program, City of Hope, Duarte, CA
- Integrated Islet Distribution Program
- Integrated Islet Distribution Program and Human Islet Phenotyping Program
Protocol Citation: Integrated Islet Distribution Program 2020. Packaging and Cold Shipping of Human Islets. protocols.io https://dx.doi.org/10.17504/protocols.io.bhdzj276
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: June 10, 2020
Last Modified: June 11, 2020
Protocol Integer ID: 38041
Keywords: Cold Shipping, TipTemp, Prodo, Human Islets, Integrated Islet Distribution Center, IIDP, Dithizone, Purity
Abstract
This SOP defines a standardized method for packaging and cold shipping of research quality islets to approved investigators of human isolated islet preparations, for use in the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) sponsored research in the Integrated Islet Distribution Program (IIDP). This protocol is written to assist the participating islet isolation centers and investigators who are part of this program.
Note
Integrated Islet Distribution Program (IIDP) (RRID:SCR_014387)
Note: This SOP was developed based on the Prodo Labs, Inc. shipping protocol and results from preliminary studies conducted by the IIDP and commissioned by the original IIDP Project Officer, and External Evaluation Committee (EEC).
Guidelines
Responsibilities:
- It is the responsibility of the IIDP CC to both follow and ensure adherence to the procedures outlined in this SOP. In order to accomplish this, the IIDP CC will interact with the relevant personnel from each of the participating centers.
- It is the responsibility of each IIDP center to follow the procedures listed in this SOP and to work to the best of their ability to follow all requirements.
Definitions:
- Integrated Islet Distribution Program (IIDP) (RRID:SCR_014387): The IIDP is a program commissioned and funded by the NIDDK to provide quality human islets to the diabetes research community to advance scientific discoveries and translational medicine. The IIDP consists of the NIDDK Project Scientist and Program Official, the External Evaluation Committee and the CC at City of Hope (COH). The IIDP CC integrates an interactive group of academic laboratories including the subcontracted IIDP centers.
- IIDP Coordinating Center (CC): Joyce Niland, Ph.D., IIDP Principal Investigator leads CC staff to coordinate the activities of the IIDP and assists the participating centers and investigators in the distribution of human islets.
- Islet Equivalent (IEQ): An IEQ is defined as an islet with a diameter of 150 microns. The number of IEQ in each size class is calculated by multiplying the number of islets by a conversion factor for each micron size.
- Approved Investigators: Researchers who have requested islets from the IIDP for basic science studies and whose research protocols have been reviewed and approved by the IIDP.
- Islet Allocation System (IA): This is the online system administered by the IIDP to allow fair distribution of basic science islets to approved investigators. This interactive System is used by the IIDP Centers and the Approved Investigators facilitates and tracks the distribution of human islets.
Materials
MATERIALS
Dynamic Diagnostics TransPak™ Pre-filled 10% Neutral Buffered Formalin (NBF)Fisher ScientificCatalog #22-899-402
Corning™ Ciprofloxacin HydrochlorideFisher ScientificCatalog #MT61277RG (Corning™ 61277RG)
Human AB Serum (ABS) HIGemini BioproductsCatalog #100-512; Heat Inactivated
PIM(G)® (5 mL Glutamine/Glutathione)Prodo Laboratories, IncCatalog #PIM(G)®
PIM(T)®Prodo Laboratories, IncCatalog # PIM(T)®
Equipment
Phase 22 PCM Flex Pack
NAME
Phase 22 cold packs to maintain temperature at 22C
TYPE
TCP Reliable Manufacturing
BRAND
FMIS76000
SKU
LINK
•Most used for applications that need to maintain a controlled room temperature
•Phase change temperature is 22°C
•Provides thermal protection when shipping products between 15-30°C
SPECIFICATIONS
Equipment
Thermo Scientific™ Nalgene™ Square PETG Media Bottles with Closure
NAME
30 mL PETG Media Bottles with Closure
TYPE
Thermo Scientific™ Nalgene™
BRAND
03-311-1V
SKU
LINK
30 mL PETG Media Bottles
SPECIFICATIONS
Equipment
PETG Media Bottles with Closure
NAME
60 mL PETG Media Bottles with Closure
TYPE
Thermo Scientific™ Nalgene™ Square PETG Media Bott
BRAND
03-311-1W
SKU
LINK
PETG Media Bottles
SPECIFICATIONS
Equipment
Media Bottles with Closure
NAME
125 mL PETG Media Bottles with Closure
TYPE
Thermo Scientific™ Nalgene™
BRAND
03-312-1
SKU
LINK
PETG Media Bottles
SPECIFICATIONS
Equipment
Durasorb™ Underpads
NAME
Underpads
TYPE
Covidien
BRAND
22-031-340
SKU
LINK
Wings™ Fluff Underpad 17 x 24 in.
SPECIFICATIONS
Equipment
Reclosable Zip Bags
NAME
Clear Reclosable Zip Bags
TYPE
RD Plastics
BRAND
19-130-6043
SKU
LINK
PE (Polyethylene) Reclosable bag (8 x 10 in.)
SPECIFICATIONS
Equipment
Uline Corrugated Boxes
NAME
Corrugated Boxes (Inner Shipping Box)
TYPE
Uline
BRAND
S-4344
SKU
LINK
7 x 5 x 5" Corrugated Boxes
SPECIFICATIONS
Equipment
Insulated Foam Shipping Kit
NAME
Insulated Foam Shipping Kit
TYPE
Uline
BRAND
S-7359
SKU
LINK
Insulated Foam Shipping Kit - 8 x 6 x 9"
SPECIFICATIONS
Equipment
Uline Cold Pack
NAME
Cold Pack
TYPE
Uline
BRAND
S-7361
SKU
LINK
Reusable, leakproof refrigerant gel stays colder than ice. Thick 4 mil poly. 6 x 4 x 3/4"
SPECIFICATIONS
Equipment
Uline Industrial Tape
NAME
Uline Industrial Tape - 2 Mil, 2" x 110 yds, Clear
TYPE
Uline
BRAND
S-423
SKU
LINK
2 Mil, 2" x 110 yds, Clear. DURABLE: Industrial Tape stands up to heat, humidity and cold.
SPECIFICATIONS
Equipment
Custom Cold Chain Complete Card
NAME
Temperature Monitoring system
TYPE
Tip TEMPerature Products
BRAND
WMSSEN044;
SKU
LINK
Custom Cold Chain Complete Card
SPECIFICATIONS
Equipment
Tip TEMPerature Products
NAME
WarmMark Ascending Temperature indicator
TYPE
WarmMark Ascending Temperature indicator
BRAND
WMSSEN009
SKU
LINK
Maximum exposure above 37°C/99°F
SPECIFICATIONS
Equipment
ColdMark Descending Temperature Indicator
NAME
Descending Temperature Indicator
TYPE
Tip TEMPerature Products
BRAND
CLMSEN004
SKU
LINK
Falling Temp Monitor Changes Color Permanently; 2°C/36°F
SPECIFICATIONS
Equipment
Laser Labels
NAME
Uline Laser Labels - White, 8 1⁄2 x 11"
TYPE
Uline
BRAND
S-5045
SKU
LINK
Box of 100 Single Labels, White, 8 1⁄2 x 11"
SPECIFICATIONS
Safety warnings
10% Neutral Buffered Formalin:
SDS_DYNDIA_FORMALIN_PREFILL_10PERCENT_NBF_40ML_20ML.pdf
SDS_DYNDIA_FORMALIN_PREFILL_10PERCENT_NBF_40ML_20ML.pdf - Hazard Statement(s): Harmful if inhaled. Causes skin irritation. Causes serious eye damage. May cause allergy or asthma symptoms or breathing difficulties if inhaled. May cause an allergic skin reaction. May cause genetic defects. May cause cancer. May cause damage to organs (lungs, nose). May cause damage to organs (lungs, nose) through prolonged or repeated exposure.
- Precautionary Statement(s): Prevention: Avoid breathing dust, vapors. Use only outdoors or in a well-ventilated area. Wash body thoroughly after handling. Wear protective gloves. Wear eye protection, face protection. Wear NIOSH approved respiratory protection. Contaminated work clothing must not be allowed out of the workplace. Obtain special instructions before use. Do not handle until all safety precautions have been read and understood. Wear protective gloves, protective clothing, eye protection and face protection. Do not breathe dust, vapors. Do not eat, drink or smoke when using this product.
Ciprofloxacin Hydrochloride SDS-61-277-AGHS-EN-2014Jul22.pdf
SDS-61-277-AGHS-EN-2014Jul22.pdf Precautionary statements:
- P280 - Wear protective gloves and eye/face protection
- P305 + P351 + P338 - IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing.
- P337 + P313 - If eye irritation persists: Get medical advice/attention.
- P273 - Avoid release to the environment.
GemCell™ U.S. Origin Human Serum ABsds_100-512_gemcell_hsab.pdf
sds_100-512_gemcell_hsab.pdf GemCell™ human serum AB is collected from healthy male donors of the AB serotype at FDA-licensed facilities in the United States.
Hazardous Components:
- Biohazard contains human source material. Handle as though capable of transmitting infectious agents.
- Toxicity: Not Established.
- Target Organs/Systems: Product could possibly irritate the skin, eyes and respiratory system. Do not ingest this product.
Before start
References:
- Prodo Labs, Inc. Protocols and Website: http://www.prodolabs.com
Prodo Media-Preparation-and-Use-Instructions-1.pdf CITATION
CITATION
Materials
Materials
The IIDP will provide each center with the supplies necessary for shipments as listed in the materials section. Standard lab supplies will be provided by the IIDP Centers as listed below.
Supplies provided by the IIDP Centers:
- Islets for distribution
- Routine lab supplies for transferring islets
- Completed FedEx Shipping Labels
- Preprinted Fresh Tissue Box Labels
- Islet Allocation Tissue Shipment Forms for appropriate recipients
- Markers for labeling vessels
Supplies provided by the IIDP:
The majority of supplies should be stored in appropriate dry, temperature-controlled environments (room temperature 16°-28°C).
- The Prodo Labs PIM(T)® should be stored, in the dark, between 2° and 8°C upon receipt but is stable at room temperature.
- The Gemini AB serum and the PIM(G)® vials should be stored at -5° to -20°C in the dark.
- The Ciprofloxacin can be stored on the shelf but filter sterilized suspension aliquots should be stored at -5° to -20°C. Aliquots:
Note
Record media preparation on Attachment 1: Solutions Preparation Sheet :
Attachment 1-Solutions preparation Sheets.pdf - The Cold Chain Complete Card with the WarmMark indicator on the card and the separate ColdMark indicator can both be stored at room temperature or between 12°C to 32°C to ensure against accidental triggering of either indicator.
- Ice Packs and CryoPaks.
Collection of Islets for Transport
Collection of Islets for Transport
After the islets have been allowed to recover from the insult of the isolation process at 37°C for approximately 18-48 hours, the quality of the cultured islets should be assessed by inspecting the islets in several flasks. If the islets have rounded up to greater than 80% of all size islets, the isolation should be ready for shipping. See attachment: Tips for Determining Islet Quality for Shipment.
Attachment 2- Tips for Determining Islet Quality for Shipment (v1).pdf Preparation of PIM(T) Media:
- Prepare one 500 mL bottle of PIM(T) media prior to the isolation
- Thaw and add 5 mL of PIM(G)
- Thaw and add 25 mL of AB serum (5% v/v)
- Thaw and add 0.5 mL of prepared ciprofloxacin sterile aliquot
- Once all additives have been added to the bottle of PIM(T), it is now referred to as PIM(T) complete.
Note
If a prepared media bottle is to be used from a previous isolation, it must have been filter sterilized at the end of the previous use. The media will expire within 30 days, once it has been fully supplemented.
Note
Record media preparation on Attachment 1: Solutions Preparation Sheet, of this SOP.
Obtain the flasks containing islets from the incubator and examine each culture flask for contamination and aseptically place inside the hood. Obtain one 250 mL conical for each 6 culture flasks and aseptically place inside hood.
Pool 6 T-175 flasks into each “settling” 250 mL conical by pipetting with a 25 mL pipette wetted with PIM(T). Repeat pooling for each set of 6 flasks into a new “settling” 250 mL conical. Allow conicals to settle while rinsing and checking the flasks.
Wash the flasks with PIM(T) to collect any remaining islets. Using a 25 mL pipette, deliver 20 mL of media slowly onto the entire bottom surface of the flask in a sweeping motion to loosen islets into the solution. Let the media run to the bottom of the flask by gravity. Repeat this step 2-3 times collecting the 20 mL from the bottom of the flask and delivering it to the top of the flask in a sweeping motion while rocking gently. The same 20 mL can be used for 6 flasks by transferring to the next flask after washing each flask. Collect the wash from all the flasks in to one “count” 250 mL conical.
Repeat wash on the next 6 flasks and place into the “count conical”.
After the first wash, examine the flasks under the microscope for stuck islets. If there are many islets attached to the bottom of flask, deliver 20 mL of PIM(T) to the flask for the second time, gently tap on the side of the flask and rock the flask back and forth to get the released islets in to the solution being careful not to get medium into the cap of the flask. Collect the wash and pool in to the “count conical”.
After the rinses and the “settling conicals” have rested for approximately 10-15 minutes, pre-wet a 25 mL pipette with PIM(T), gently draw the settled islets from the bottom of each of the “settling conicals”, one at a time, by going to the bottom of the conicals containing the settled islets, and pipette with upward movement with only ~ 2-3 mL of media and carefully deliver to the bottom of 250 mL “count conical”.
Centrifuge the supernatant in the “settling conicals” for 2 minutes @180 g.
After centrifugation, evacuate supernatant. Gently resuspend the pellets from the “settling conicals” with 2-3 mL of PIM(T) and transfer into the “count conical”.
Bring count sample up to 200 mL with PIM(T). Take count samples by inverting 250 mL conical 3-4 times, quickly remove cap and have second technician remove exactly 0.5 mL form the “count conical” and place in counting dish. Place the conical containing islets horizontally under the hood while waiting for results from the evaluations.
According to the islet counts, calculate the concentration of islets/mL and record in the batch record.
Attachment 3-Islet Tabulation Counting Sheet for Shipping Islets (v1).xlsx Complete the broadcast records and proceed to broadcast. From this value, calculate how much volume is needed to be distributed into each shipping bottle for each order. Record the value in the batch record.
Remove 100 IEQ for Viability Test and proceed according to IIDP protocol.
Remove 400 IEQ for GSIR studies and culture per IIDP protocol.
As investigator’s orders are determined through the broadcast system, prepare appropriate amounts of islets into proper sized shipping vessel.
- 30 mL bottle for shipments of less than 10,000 IEQ
- 60 mL bottle for shipments of 10,000-50,000 IEQ
- 125 mL bottle for shipments of greater than 50,000IEQ - 100,000 IEQ
Distribute appropriate volume into shipping bottles.
Bring the volume up with PIM(T) stored at 6-10ºC up to the halfway point of the neck of each shipping bottle. Note: Specific volumes for specific bottles sizes are as follows:
- 30 mL bottle – 44 mL of media plus islets
- 260 mL bottle – 75 mL of media plus islets
- 125 mL bottle – 167 mL of media plus islets
Label each bottle with RRID #, Date, #IEQ, and PI Name
Record appropriate information on the batch record.
Keep the shipping bottles with islets in the refrigerator set at 6-10º C until ready for packaging.
Packaging of Islets
Packaging of Islets
Stage all materials needed for packaging the islets and ensure all ice packs and CryoPak
pouches are at the proper temperature.
The Cold Chain Complete Card is shipped with the WarmMark Indicator attached.
- Store the Card at least 5ºC (9º F) below the activation temperature of the adhered WarmMark of 37ᵒC (therefore less than 32º C). Room temperature is acceptable.
- The ColdMark is provided separately because it is always active and should be stored at least 10ᵒC (50ºF) above activation temperature of ColdMark of 2ºC (therefore more than 12ºC).Room temperature is acceptable.
- When ready to use, adhere the ColdMark indicator to the card and activate the WarmMark. ACTIVATE the WarmMark by folding up and pulling out the tab until completely removed.
- Immediately place the Cold Chain Complete Card along with the islet shipping bottles into a Ziploc bag and seal.
- Include attached Cold Chain Complete Form
Attachment 4 -TipTemp Handling Instructions (v1).pdf Line the inner cardboard (S-4434) box with absorbent pad. See attached slide presentation for more information.
Attachment 5-Packaging Islets for Cold Shipping.pdf - Place the bagged shipping bottle(s) containing the islets in the Ziploc bag into the absorbent pad lined, inner cardboard box.
- Surround bottle with six 8ºC cold packs, one on bottom, one on each of the four sides and one on top.
Mark the direction for upright bottle. Seal the inner box with shipping tape and stand upright.
Place one 8°C CryoPak pouch and one 8°C cold pack (stored in the refrigerator) in the bottom of the Styrofoam box.
Place the inner cardboard box containing the islets on top of the pouch and pack, ensuring that the bottle inside is standing upright and centered in the shipping container.
Place one 8°C CryoPak pouch and one -20°C frozen ice pack on each long side of the inner box. (Packs/pouches are placed in the larger gap between the inner box and the insulated shipping container.)
Place one 8°C CryoPak pouch and one 8°C cold pack on the top of the inner box. *NOTE-All packs should be placed in an alternating method, to ensure two of the same packs are not next to each other in the configuration (See Attachment 5-Packaging Islets for Cold Shipping for visual details.)
Place the Styrofoam lid on top.
Place Tissue Shipment Form on top of Styrofoam lid.
Close the flaps of the shipping box and seal with shipping tape. Add Box label to the Box.
Attachment 6-Generic Institution Box Label IIDP 2019 (v1).pdf Attach FedEx label with recipient FedEx number to box and ship priority overnight.
Completing the Distribution Records
Completing the Distribution Records
The shipping center will enter the IEQ to be shipped (Confirm Recipient screen) and will enter FedEx tracking numbers (Islet Tracking) into the Islet Allocation System. In addition, the shipping center should process the shipment by using the on-line FedEx process and click the prompt to alert the recipient of the FedEx tracking number. This will help the recipient investigator follow their shipment through the FedEx system and free up time for the centers and the IIDP.
Note: If the investigator has picked up the islets at the distributing center then the center should check “Yes” in the “Direct Pickup” column and “Yes” under “Islets Shipped?” once they are picked up.
As soon as the center has entered the number of IEQ shipped and indicated that the investigator will be shipped islets, the investigator’s account will be set automatically to “ineligible”. This will act similar to “on hold” preventing the investigator from receiving additional targeted offers until the User Feedback Form has been completed.
The investigators will have the FedEx tracking numbers listed in their History of Islet Offers table.
Citations
Ricordi C, Gray DWR, Hering BJ, Kaufman DB, Warnek GL, Kneteman N, et al.. Islet isolation assessment in man and large animals.
http://www.ncbi.nlm.nih.gov/pubmed/2075782David W. Scharp, Jayagowri Arulmoli, Kelly Morgan, Hannah Sunshine, Ergeng Hao. Advances in Human Islet Processing: Manufacturing Steps to Achieve Predictable Islet Outcomes from Research Pancreases
10.21926/obm.transplant.1901052