GUIDELINES FOR TRANSPORT OF
FROZEN GAMETES AND EMBRYOS
ART Laboratory Special Interest Group (ART Lab SIG)
Working Group, ART Laboratory Special Interest Group
Chair: Haimant Bissessar, BSc
Belen Herrero, PhD
Lynda Hughes, BSc
Murid Javed, PhD
David Mortimer, PhD
Jill Mellon, MSc
Peter Przybylski, MSc
April 2016
This document is based on available evidence and practices to date, often in a rapidly advancing field.
Recommendations may not reflect emerging technologies or practices and are subject to change.
Guidelines are intended as an aid to practice, and do not dictate an exclusive course of action.
All rights reserved © 2016 Canadian Fertility and Andrology Society. This document may not be
reproduced in its entirety or in part without the expressed written consent of the Canadian Fertility and
Andrology Society.
Table of Contents
Introduction:
3
Section 1: Transport process
4
Section 2: Informative Documents
5
Section 3: Warming solutions (kits)
6
Section 4: Information about vapour (dry) shippers
6
4.1. Vapour (dry) shipper preparation
7
4.2. Receiving dry shipper/receiving vitrified gametes or embryos
8
4.3. Vapour (dry) shipper decontamination
8
Section 5: Common problems encountered during transport procedure
9
5.1 Potential operator errors.
9
5.2 Potential cryogenic shipping tank problems.
9
5.3 Potential transportation problems.
10
Appendix 1: Flowchart for the transport of human gametes and embryos
11
Appendix 2: List of Companies which specialize in the transport of
cryopreserved gametes and embryos, as of December 2015*
12
Appendix 3: List of companies which supply exposure indicators
and dataloggers, as of December 2015*
13
Appendix 4: Sample checklist for the transportation of
cryopreserved material between ART clinics.
14
References:
15
Guidelines for Transport of Frozen Gametes and Embryos – Published May 17, 2016
2
Introduction:
For decades the transport of frozen gametes and embryos has occurred between cryobanks,
fertility clinics and cryo repositories. The risks to gamete and embryo integrity and potential loss
due to vapour (dry) shipper failure, improper handling and temperature fluctuations inside the
vapour (dry) shipper have always been present. These risks are significantly increased with
gametes and embryos cryopreserved by vitrification since vitrified gametes and embryos are
stored in a very small volume of medium making them far more susceptible to inadvertent
warming.
The need for national transport guidelines was raised at the ART Lab SIG meeting during the 2014
CFAS Annual Meeting in Quebec City. The ART Lab SIG accepted the task and decided to create
a working group to develop the guidelines. A call for volunteers was initiated by a general e-mail
to all ART Lab SIG members and the working group was created.
The following guidelines have been developed to provide guidance to Andrology and Embryology
laboratory personnel and patients on current best practice for the transfer of gametes and
embryos from one location to another. They are not intended to be a protocol to be applied in
all situations.
It is our hope that ART professionals and patients will benefit from these guidelines.
Haimant Bissessar
Working Group Chair
Guidelines for Transport of Frozen Gametes and Embryos – Published May 17, 2016
3
Section 1: Transport process
1.1. Frozen donor or patient samples must be transported using a vapour (dry) shipper which
has been designed specifically for this purpose, holding samples at temperatures of –
150°C or colder. Transport on dry ice (–78.5°C) must be avoided as this can compromise
the quality of the frozen material since it is above the ice transition point of water (ca. –
132°C). A Flowchart for the transport of human gametes and embryos can be found in
Appendix 1.
1.2. It is recommended to use a courier company which specializes in the transportation of
cryopreserved biological specimens as there are potential risks associated with the
transport. Patients should contact the courier company for risk assessment. A list of
companies can be found in Appendix 2.
1.3. The shipping and receiving laboratories must arrange a date and time that is mutually
convenient to facilitate the transport.
1.4. Laboratory personnel must ensure that samples are accompanied by all relevant
paperwork including permits and customs declarations for international shipments
(Refer to section 2).
1.5. Since the logistics of organizing the transportation can take time, patients should be
advised to allow sufficient time when making the arrangements so that their treatment
is not delayed.
1.6. When transporting a patient’s stored material, centres must inform patients of the risks
associated with vapour shipper failure and the risks inherent in transferring all of the
patient’s samples in a single shipment (Refer to item 1.7).
1.7. Operator and tank complications can be minimized by developing, implementing and
following proper Standard Operating Procedures (SOPs) and Quality Assurance (QA)
protocols. Only trained and experienced personnel should maintain the vapour shippers
and ship gametes and embryos. Use of thermal exposure indicators or dataloggers which
keep a history of the temperature inside the shipper during the shipping process is
recommended. A list of suppliers of exposure indicators and dataloggers can be found in
Appendix 3.
Guidelines for Transport of Frozen Gametes and Embryos– Published May 17, 2016
4
Section 2: Informative Documents
Centres must ensure that all documents are in place when proceeding with the transport process
and have a documented training program for all personnel involved in the process.
2.1. Centres must provide comprehensive information regarding the transferred material
including but not limited to the following: - See sample checklist in Appendix 4.
a) Cryopreservation date, quantity and quality of samples transferred.
b) Information
on
the
cryopreservation
method
(vitrification/slow
freezing),
cryocontainer and number of oocytes or embryos in each cryocontainer or details of
the cryopreserved sperm.
c) A detailed description of the thawing method to be performed.
d) Serology testing results performed at the time the gametes/embryos were frozen and
any other pertinent information including post quarantine serology results if
applicable. Serology testing may be mandated as in the Health Canada Semen
Regulations or considered best practice as determine by industry standards and
practice guidelines. Any positive or missing serology results may prohibit shipping of
gametes/embryos.
e) If completed, the consent to use as defined under section 8 of the Assisted Human
Reproduction Act.
2.2. A process must be in place to allow patients to take responsibility for the transport of their
own specimens. Before proceeding with a transport, the centres must request the
owner(s) of the gametes or embryos to sign consent to transport agreement. Photo
identification from the owner(s) of the gametes or embryos should be requested and a
copy filed in the owner’s clinical records.
2.3. At the receiving facility, documentation should be provided to patients regarding the
receipt and storage of samples. A storage agreement should be signed by the owner(s) of
the frozen material. Photo identification from the owner(s) of the gametes or embryos
must be requested at all times and a copy filed with the owner’s clinical records.
Guidelines for Transport of Frozen Gametes and Embryos– Published May 17, 2016
5
Section 3: Warming solutions (kits)
3.1 In addition to information about the number and quantity of gametes or embryos
cryopreserved, in the case of vitrified gametes or embryos, the vitrification system devices
and instructions on warming must be provided with the shipment.
3.2 The use of warming kits that are different from the kit or system used to vitrify the
specimen(s) constitutes “off label” use in terms of product regulatory approvals and is not
recommended. Patients must be advised of the risk associated with such off-label use of
warming kits.
Section 4: Information about vapour (dry) shippers
There are different manufacturers, suppliers and types of vapour shippers. Vapour shippers are
manufactured from durable, lightweight aluminum with a hydrophobic compound on the inside
that absorbs liquid nitrogen. This compound allows for dry, spill free vapour-phase shipping. The
shipper is called a “dry” shipper because it no longer contains any free liquid nitrogen. Vapour
shippers are manufactured with varying hold times, with some shippers designed to maintain a
minimum internal temperature of -150oC for up to 3 weeks. If the shipper is shipped on its side
it will only provide 40% of the specified hold time. If shipped upside down it will only provide
10% of the hold time capacity.1 A properly prepared dry shipper is not considered to be
“hazardous goods” under International Air Transport Association (IATA) regulations.2 The words
“Not Restricted as per Special Provisions A152” , “Fragile Human Specimens”, DO NOT OPEN,
must be clearly indicated on the air way bill and all documentation under the International Civil
Aviation Organization (ICAO) regulations.3 While a properly prepared vapour shipper does not
contain free liquid nitrogen it does contain ultra-cold nitrogen vapour. If liquid nitrogen is visible
in the bottom of the dry shipper at the time of shipment it is not eligible for shipment by air. 4,5
Guidelines for Transport of Frozen Gametes and Embryos– Published May 17, 2016
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4.1. Vapour (dry) shipper preparation
Caution: Liquid nitrogen evaporates to give nearly 700 volumes of nitrogen gas making it a simple
asphyxiant. Consequently it must only be used in a well-ventilated room equipped with an
oxygen depletion sensor and alarms. All personnel must receive adequate training in liquid
nitrogen handling and use and must never work alone. Manufacturer’s instructions for filling
vapour shippers must be followed. Refer to the Material Safety Data Sheet (MSDS)
documentation before use. It is highly recommended that two individuals be involved in the
loading and receiving of gametes and/or embryos. Document and sign off on each step in the
process.
1. Position the empty vapour shipper in a location out of the traffic flow where it will not be
disturbed. Weigh and document weight.
2. Ensure that appropriate personal protective equipment is worn when working with liquid
nitrogen: (i.e. insulated gloves, thermal apron, fully enclosed shoes and a face shield).
3. Place the shipper on the floor and fill slowly with liquid nitrogen from a supply tank.
4. Allow the nitrogen to gradually lower the temperature of the dry shipper. This will
require 15 to 20 minutes, depending upon the capacity of the supply tank dispensing hose
and condition of the dry shipper. Up to 20 litres of liquid nitrogen can be required to fill
a typical vapour shipper.
5. A significant amount of nitrogen gas will be generated as the cold liquid contacts the
warm surfaces inside the shipper. Therefore, always add the liquid nitrogen slowly and
work in a well-ventilated space.
6. Pouring liquid nitrogen into a warm or room-temperature dry shipper too quickly will
cause the nitrogen to “boil over” or splash out the top of the dry shipper, possibly
splashing the user and causing skin burns.
7. When the liquid reaches the neck of the vapour shipper, stop filling. Replace the cap and
set the dry shipper aside for the period specified by the manufacturer to allow the liquid
nitrogen to saturate the absorbent material inside the tank.
8. Repeat these steps until the liquid level no longer decreases on standing. This may require
as many as 15 repetitions over 24 hours.
Guidelines for Transport of Frozen Gametes and Embryos– Published May 17, 2016
7
9. Remove all free liquid nitrogen from the vapour shipper before transport. Empty the dry
shipper by pouring the excess liquid nitrogen back into a large liquid nitrogen Dewar.
10. Weigh the dry shipper prior to loading with gametes or embryos. Document weight.
11. Advise the transport company to always keep the dry shipper in an upright position. This
can be accomplished by affixing directional arrows to the outside of the shipping
container in addition to labels to indicate “DO NOT OPEN”, Fragile, Human Specimens.
4.2. Receiving dry shipper/receiving vitrified gametes or embryos
1. Inspect the outer shipping container for signs of physical damage.
2. Open outer shipping container to expose vapor shipper and inspect for damage.
3. Remove and review shipping documents prior to opening of shipper.
4. Remove security seal (if applicable) and remove lid of shipper. A “puff” of liquid nitrogen
vapour should be seen indicating that the shipper has maintained cryogenic temperature.
However the use of data loggers or other temperature measuring devices is
recommended. For vitrified material the shipper must be re-filled with liquid nitrogen.
Do not stick the end of the supply hose into the shipper and then dispense liquid nitrogen
as the vitrified material will be subject to a blast of warm air from the hose. Liquid
nitrogen must be dispensed into a container approved for liquid nitrogen use and then
pour into shipper.
4.3. Vapour (dry) shipper decontamination
Sanitizing and Decontaminating MVE aluminum vapour shippers
1. Any cleaning solution that does not react with aluminum or stainless steel can be used in
the sanitation process.
2. In most cases many household detergent or mild soap solutions may be suitable or a
solution of 10% chlorine bleach and 90% water.
3. Allow the internal temperature of the dry shipper to warm to room temperature.
4. Spray the outside and completely fill the vapour shipper with cleaning solution and leave
for 30 minutes.
Guidelines for Transport of Frozen Gametes and Embryos– Published May 17, 2016
8
5. Decant the solution and rinse the shipper with clean water.
6. Place the shipper on its end (inverted) and allow to completely dry before allowing into
service.
NOTE: DO NOT USE ANY PETROLEUM BASED CLEANING SOLUTION.
Section 5: Common problems encountered during transport procedure
5.1 Potential operator errors.
1. Improper cooling of a dry shipper tank.
2. Improper maintenance/quality assurance of a dry shipper tank.
3. Improper transfer of specimens from the cryostorage tank to the dry shipper, and vice
versa.
4. Sending the wrong gametes or embryos and/or the incorrect number of gametes or
embryos.
5. Shipping to the wrong address.
6. Incomplete shipping documentation (different for domestic vs international).
7. Not recording dry shipper condition and temperature when shipping and receiving tanks.
5.2 Potential cryogenic shipping tank problems.
1. Loss of dry shipper tank vacuum due to physical damage to the tank.
2. Damage to the liquid nitrogen adsorbent material inside the vapour shipper.
3. Mechanical damage.
4. Inadequate temperature inside the vapour shipper.
5. Age of the shipper.
6. Inadequate/missing shipping labels (address, non-biohazard, etc.).
Guidelines for Transport of Frozen Gametes and Embryos– Published May 17, 2016
9
5.3 Potential transportation problems.
1. Shipping delays due to mechanical (truck, train, or plane) problems.
2. Mis-sorting or mis-direction by courier (sent on wrong truck, train, or plane).
3. Delayed pickup and/or delivery.
4. Customs delays in the case of international shipping.
5. Adverse weather conditions.
6. Mishandling of the shipper (e.g. not being kept upright, tipping, falling/dropping from
cargo bays, vehicles, etc.).
7. Vandalism.
8. Potential exposure of embryos by airline/customs personnel opening and inspecting the
contents. This can be avoided by using a padlock: a Transportation Security
Administration (TSA) approved combination padlock should be used so that customs
personnel can open it if necessary, but this will be evident until the user re-sets the lock.
Guidelines for Transport of Frozen Gametes and Embryos– Published May 17, 2016
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Appendix 1: Flowchart for the transport of human gametes and embryos
Patients contact receiving clinic with
their request to transport their gametes
or embryos
FROM YOUR FACILITY TO ANOTHER
FROM ANOTHER FACILITY TO YOURS
Patient is provided with a laboratory
contact at your clinic once their
records have been transferred
Patients arrange consult at your clinic
and transfer their record
Receiving lab makes contact to
determine suitability of specimens for
transport
Your lab contacts the facility where
specimens are stored to determine
their suitability for transport
OK TO TRANSPORT
NOT SUITABLE FOR
TRANSPORT
USING COURIER
PATIENTS USING
CLINIC SHIPPER
Patients are
given a list of
couriers
specializing in
shipping of
gametes and
embryos
The receiving lab
provides shipper
and contacts your
lab to arrange a
convenient time
to send their
shipper
The recipient
lab contacts
patients with
reason why
specimens
can't be
transported
OK TO TRANSPORT
USING COURIER
Patients arrange
courier. You will
be informed by
the courier as to
when to expect
the shipment to
take place
Guidelines for Transport of Frozen Gametes and Embryos – Published May 17, 2016
NOT SUITABLE FOR
TRANSPORT
PATIENTS USING
CLINIC SHIPPER
Your lab
contacts
patients
with reason
why
specimens
can't be
transported
Your lab provides
shipper and
contacts the
facility where
specimens are
stored to arrange
a convenient
shipping time
11
Appendix 2: List of Companies which specialize in the transport of
cryopreserved gametes and embryos, as of December 2015*
Company
Website address
Phone number
e-mail
Area served
Axis Global
http://www.axisg.com/
1-800 568-4901
[email protected]
International
Core Cryo Lab
www.corecryolab.com
1-866-580-9872
[email protected]
International
Corion Fertility
Clinic
Cryoport
www.corionfertilityclinic.com
01191-222-6395588 or 5599
1-866-844-5150
drkadam@corionfertilit
yclinic.com
[email protected]
India
DHL
http://international.dhl.ca/en/logi 1-855-345-7447
stics/customer_resource_area/logi
stics_news/spotlight_newsletter/0
8132014.html
Contact form can be
completed from
website
International
KynisiCourier
www.kynisi.com
[email protected]
International
Marken Express
www.marken.com
01144-207-3570001
1-800-627-5361
[email protected]
U.S./Canada
Medair Time
Critical Express
Outreach Health
Services
Repromed Ltd.
www.medair.ca
1-905-507-4780
[email protected]
Canada
www.creatingcanadianfamilies.ca
1-866-785-4709
International
www.repromed.ca
1-877-317-6079
TT Courier
www.ttcourier.in
UPS
https://www.ups.com/content/ca
/en/shipping/ship/premiumcare.html?srch_pos=1&srch_phr=
premium+care
01191-222-8328374 or 8375
1-800-PICK-UPS
(ask to speak to
Account
Manager)
info@creatingcanadianf
amilies.ca
patientservices@repro
med.ca
[email protected]
World Courier
www.worldcourier.com
1-604-232-9444
1-905-362-9462
Xytex Tissues
www.xytextissues.com
1-800-277-3210
www.cryoport.com
International
Canada
India
International
International
[email protected]
U.S./Canada
*The above list of companies is not exhaustive as there are other companies that transport gametes and
embryos. The companies have not been evaluated nor endorsed by CFAS.
Guidelines for Transport of Frozen Gametes and Embryos – Published May 17, 2016
12
Appendix 3: List of companies which supply exposure indicators and
dataloggers, as of December 2015*
Company
Product
Website address
Phone number
e-mail
Cryoguard
Corporation
Thermal
exposure
indicators
http://www.cryoguard.com
http://www.cryoguard.com/m1201/
215-712-9018
customer_service@cryo
guard.com
ELPRO
Services Inc.
LIBERO PDF
dataloggers
www.elpro.com
740-568-9900
[email protected]
Chart-Biomed
MVE
dataloggers
http://files.chartindustries.com/
ML111006%20A18.1baccessories%20catalog%20.pdf
1-800-482-2473
MVE.request@chartind
ustries.com
+44 1932 755000
[email protected]
http://www.chartindustries.com
/Life-Sciences/Parts-andAccessories
Planer PLC
ShipsLog
Datalogger
https://planer.com/products/da
ta-logging.html
*The above list of companies is not exhaustive as there are other companies that transport gametes and
embryos. The companies have not been evaluated nor endorsed by CFAS.
Guidelines for Transport of Frozen Gametes and Embryos– Published May 17, 2016
13
Appendix 4 Sample checklist for the transportation of cryopreserved material
between ART clinics
Patient
Partner
Sender Clinic
____________________________________
Recipient Clinic
____________________________________
Type of material
SPERM
EGGS
EMBRYOS
If embryos, which day of development _______________
Laboratory resource person at sender clinic
Name:
Direct tel. number:
Email:
Media used for cryopreservation:
Device used for cryopreservation:
Instructions included:
YES
NO
Important information about warming:
Embryology report included
Shipper filled and excess liquid nitrogen removed before shipping
Consent for original use
Source: 2015 meeting of Montreal area IVF clinics sponsored by Merck Canada.
Guidelines for Transport of Frozen Gametes and Embryos – Published May 17, 2016
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References:
1
MVE Operating Instruction Manual for Vapor Shipper Dewars
Chart Industries, Inc.
Chart MVE Biomedical Division,
2200 Airport Industrial Dr. Suite 500; Ball Ground, GA 30107 USA
www.chartbiomed.com Phone number: 770-721-7759
2
International Air Transportation Association (IATA) Dangerous Goods Regulation 2.3.4.3
http://www.iata.com
3
International Civil Aviation Organization (ICAO) Doc 9284 — Technical Instructions for the Safe
Transport of Dangerous Goods by Air.
http://www.icao.int/Search/pages/Results.aspx?k=liquid%20nitrogen%20shippers
4
International Civil Aviation Organization (ICAO) Doc 9284 — Technical Instructions for the Safe
Transport of Dangerous Goods by Air.
http://www.icao.int/safety/DangerousGoods/DGP%2022%20Working%20Papers/DGP.22.WP.041.2.en.pdf
5 US
Code of Federal Regulations -CFR 173.320(a) Title: 49
http://www.gpo.gov/fdsys/pkg/CFR-2011-title49-vol2/xml/CFR-2011-title49-vol2-sec173-320.xml
Guidelines for Transport of Frozen Gametes and Embryos– Published May 17, 2016
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