1. Meeting medical device standards with a closed
system cryogenic vial for cellular therapy
products.
Presenter: Evonne R. Fearnot, MS
July 29, 2013, Bethesda, MD

2. Overview
Objectives for this topic are to:
– Present elementary information related to the FDA
510(k) process
– Expand understanding of the risk management
process to construct an appropriate testing strategy
– Examine a cryogenic storage vial which has made it
through such a testing strategy

3. • Medical devices fall into three classes
• With some exceptions;
Medical Device Categories
Class 1 Class 2 Class 3
Low Risk Medium Risk High Risk
Requires Device
Registration
Requires
510(k)
Requires
PMA

4. 510(k) Process
A 510(k) is required:
• When introducing a device to the market with a
predicate device similar in class for the first
time;
• A change in indications for use for a previously
cleared device;
• And if a significant modification is made to a
previously cleared device.

5. Changing Regulatory Environment
A few examples that show even 510(k) policies are
being revisited include:
• In December 2012, FDA released a guidance
document titled Refuse to Accept Policy for
510(k)s.
• The guidance document was initiated to help
timeliness of review.
• This past June 2013, FDA had a meeting titled
510(k) Device Modifications: Deciding When to
Submit a 510(k) for a Change to an Existing
Device.
• This is the first step towards the Agency initiating
a new draft guidance.

6. 510(k) Process
Fortunately today, the cryogenic storage vial to be
discussed requires a 510(k) because it is a device
being:
• Introduced to the market with a predicate device
similar in class for the first time;
• A change in indications for use for a previously
cleared device;
• And when a significant modification is made to a
previously cleared device.
Thus, for this situation, in order to obtain FDA
510(k) clearance, a device must be determined
substantially equivalent in comparison with a
predicate.

7. 510(k) Process
A device is considered substantially equivalent if, in
comparison to a predicate, it:
• has the same intended use as the predicate; and
has the same technological characteristics as the
predicate;
or
• has the same intended use as the predicate; and
has different technological characteristics and the
information submitted to FDA:
– does not raise new questions of safety and
effectiveness; and
– demonstrates that the device is at least as safe and
effective as the predicate device.

8. Case Study
The subject of the case study is this CellSeal
Closed System Cryogenic Vial.

9. Testing Is Risk Based
• Testing is based on anticipated risk.
• The identified risks include:
– Safety of patient contacting materials
– Container closure integrity
– Stability of the cell therapy product over time

10. Safety Evaluation Program of Patient
Contacting Materials
• To address the first identified risk, safety of patient
contacting materials, the Blue Book Memorandum, G95-
1, and ISO 10993-1 were used to determine an
appropriate device category and develop a satisfactory
safety evaluation program.
• The FDA and ISO 10993-1 classifies CellSeal Vial with:
– Device Type
• External communicating device
– Body Contact
• Blood path, indirect
– Contact Duration
• Permanent (>30 days)

11. Safety Evaluation Program of Patient
Contacting Materials
• This device classification essentially means the
material being stored inside the vial is being
deemed an extension of the patient, resulting in the
vial becoming a permanent patient-contacting
medical device.
• It is interesting the FDA classifies the device this
way; as designed the device may never be in the
same room as a patient and also the sample may
only be in contact with the vial for up to 24 hours
before freezing.

12. Safety Evaluation Program of Patient
Contacting Materials
• Through consideration and direct
correspondence with FDA, a safety evaluation
program was determined to include:
– Applicable biological testing, including cytotoxicity,
sensitization, irritation, pyrogenicity, hemolysis,
acute systemic toxicity, and a genotoxicity test panel,
– An extractables study with at least one extract
containing DMSO,
– And a risk assessment showing safety of the device
when used according to the manufacturer’s
instructions

13. Biological Evaluation Results
Biological Effect Test Results
Cytotoxicity ISO Neutral Red
Neat extract viability
103%. Not cytotoxic.
Sensitization ISO Kligman Maximization Test
Not a sensitizing
agent.
Irritation ISO Intracutaneous Test
No significant
irritation.
Systemic toxicity
ISO Acute Systemic No effect.
ISO Material Mediated Pyrogen Not pyrogenic.
Genotoxicity
ISO Ames Test Not mutagenic.
ISO Mouse Micronucleus Assay Not mutagenic.
ISO In Vivo Mouse Lymphoma Fwd Mutation Non-clastogenic.
Hemocompatibility
ISO Hemolysis- ASTM Direct And Indirect Contact Not hemolytic.
ISO In Vitro Hemocompatibility Assay-Direct Contact
No effect on
hematogram.
ISO Unactivated Partial Thromboplastin Time-Direct
And Indirect Contact
No effect on clotting
time.

14. Chemical Characterization of Materials
• Examination of extractables is growing in
importance especially for polymer-derived
components
• Extractables are compounds that can be
extracted from the container closure system
when in the presence of a solvent
• The extractables study used on the CellSeal
Vial included direct analysis, and exposure to
two extract model solvents
• Extraction conditions were determined to
mimic clinical use

15. Upon review of extractables data it was
revealed:
• While a number of unidentifiable compounds
were present, identified compounds detected
were considered either benign or present at
low levels.
• Due to low risk associated with identified
compounds, a leachables study was deemed
unnecessary.
Chemical Characterization of Materials

16. Risk Assessment Results
The final risk assessment revealed:
• High safety margins for identified compounds
• So, no elements or compounds that were present
pose a risk of adverse effects in the clinic
• Unidentified compounds were determined
unlikely to pose a toxicological hazard with use of
the device because of absence of effects in
biocompatibility testing
Thus, the final risk assessment supported safety of
the device materials for their intended use

17. Container Closure Integrity
The next identified risk to address is container
closure integrity.

18. • Obviously, the packaging for the device must
maintain integrity and expiry date of the shelf life
of the device prior to use.
• ISO 11607, Packaging for terminally sterilized
medical devices was used to construct a testing
plan to validate packaging integrity and shelf life
of the device.
• An acceptable and statistically significant sample
size was chosen according to product’s identified
risk category.
• And appropriate closure integrity tests were
chosen as endpoints.
Container Closure Integrity Prior to Use

19. • For packaging validation, the worst-case shipping
configuration was used.
• The worst-case shipping configuration was the
shipper that received the most transit, was the
heaviest, bulkiest, and thus, will experience the
most damage, especially to those vials on the
bottom of the shipper.
Container Closure Integrity Prior to Use

20. Container Closure Integrity Prior to Use

21. • The Environmental Conditioning sequence included
varying temperature and humidity to address both
summer and winter conditions.
Total Duration = 150 hrs
• 60°C is higher than the world’s hottest temperature.
Container Closure Integrity Prior to Use

22. • The Distribution Simulation sequence included subjecting
packaging to a first drop test from a height of 24 inches on
all of its sides.
• Vehicle Stacking Load Compression Test
Container Closure Integrity Prior to Use

23. • Vehicle Vibration Test
• Concentrated Impact Test
Container Closure Integrity Prior to Use

24. • Loose Load Vibration Test
• Low Pressure (High Altitude) Hazard Test
Container Closure Integrity Prior to Use

25. • And finally to a second drop test from a height of 24 inches
on all of its sides again and an added height of 48 inches
Container Closure Integrity Prior to Use

26. • A 3 year shelf life was selected because the predicate
device had a 3 year shelf life.
• The shelf life was validated with an accelerated aging
study followed by real-time data.
Container Closure Integrity Prior to Use

27. • As illustrated on the flow chart, product tests were
performed after subjecting the device packaging to
hazardous situations and aging. These tests included:
– Microbial Aerosol Challenge, which determined if
the device acted as an effective microbial
barrier
– and Leak Test, which determined if all the bonds
and connections of the device maintained
integrity
• All products in the package validation testing met the
requirements of the tests.
Container Closure Integrity Prior to Use

28. Container Closure Integrity During
Actual Use
• Container integrity of the device during actual use
was determined by subjecting the device to control-
rate freezing, storage in liquid nitrogen, 1-meter drop
test, transportation, and thawing.
• Following thawing, a series of container integrity
tests, which were dye and bacterial immersion, were
performed.
• All products met the requirements of both tests.

29. Stability
• Last identified risk to address includes stability
of the cell based product over time during
storage.

30. Stability
• The stability of the CellSeal Vial was compared
to the Pall Medical Freezing Bag (predicate)
through cryostorage of human umbilical cord
blood hematopoietic progenitor cells (UCB).
• 6 pre-defined time points for storage were
used (1wk, 1 mo, 1 yr, 5 yr, 10 yr, 15 yr).
• 3 donors per time point were used.

31. • For each donor, a 30 mL concentrated UCB was
prepared with a final concentration of 10% DMSO.
• 10 mL UCB was cryopreserved in two 5 mL CellSeal Vials
and the remaining 20 mL UCB was cryopreserved in one
Pall Medical Freezing Bag.
Stability
10 mL
20 mL
UCB Donor
Unit

32. Stability
• All samples were analyzed for total nucleated cell count,
colony-forming unit (CFU) assay and viability.
• Clonogenic properties of post-thawed human UCB
cryopreserved for the first time point are described in
the following table.

33. Stability Results
There was no significant difference (student’s t-test, p ≤
0.05) in the performance of the CellSeal Vial when
compared to the Pall Medical Freezing Bag, which was also
true for the segments on respective devices.
All values are indicated as mean ± SEM.

34. Case Study Conclusion
These studies confirm that the CellSeal Closed
System Cryogenic Vial meets medical device
testing standards for low volume
cryopreservation of cell based products.

35. Summary
• Understanding submission requirements is
challenging.
• Thus, planning, conduct, and documentation of
testing should be in accordance with submission
requirements and device-specific hazards.
• Finally, while freezing and safety of the sample are
critical design requirements of a cell therapy
cryogenic storage vial intuitively leading to very
specific testing, it proved necessary to greatly
extend the testing scope to considerations such as
transportation and storage, to meet medical
device standards.

36. Thank you for your attention.
Questions?
Evonne R. Fearnot, MS
evonne.fearnot@cookgbt.com