HPC (Hematopoietic Progenitor Cell), Cord Blood, is an allogeneic cord blood hematopoietic progenitor cell therapy indicated for use in unrelated donor hematopoietic progenitor cell transplantation procedures in conjunction with an appropriate preparative regimen for hematopoietic and immunologic reconstitution in patients with disorders affecting the hematopoietic system that are inherited, acquired, or result from myeloablative treatment.

The risk benefit assessment for an individual patient depends on the patient characteristics, including disease, stage, risk factors, and specific manifestations of the disease, on characteristics of the graft, and other available treatments or types of hematopoietic progenitor cells.

• For intravenous use only.
• Do not irradiate.

Unit selection and administration of HPC, Cord Blood, should be done under the direction of a physician experienced in hematopoietic progenitor cell transplantation.

Each HPC, Cord Blood unit contains a minimum of 5.0 x 108 total nucleated cells with a minimum of 1.25 x 106 viable CD34+ cells, suspended in 10% dimethyl sulfoxide (DMSO) and 1% Dextran 40, at the time of cryopreservation.

The exact pre-cryopreservation nucleated cell content is provided on the container label and in accompanying records.

None

Day-100 mortality from all causes was 25%.

The most common infusion-related adverse reactions (≥ 5%) are hypertension, vomiting, nausea, bradycardia, and fever.

HPC, Cord Blood consists of hematopoietic progenitor cells, monocytes, lymphocytes, and granulocytes from human cord blood for intravenous infusion. Blood recovered from umbilical cord and placenta is volume reduced and partially depleted of red blood cells and plasma.

The active ingredient is hematopoietic progenitor cells which express the cell surface marker CD34. The potency of the cord blood is determined by measuring the numbers of total nucleated cells (TNC) and CD34+ cells, and cell viability. Each unit of HPC, Cord Blood contains a minimum of 5 x 108 total nucleated cells with at least 1.25 x 106 viable CD34+ cells at the time of cryopreservation. The cellular composition of HPC, Cord Blood depends on the composition of cells in the blood recovered from the umbilical cord and placenta of the donor. The actual nucleated cell count, the CD34+ cell count, the ABO group, and the HLA typing are listed on the container label and/or accompanying records sent with each individual unit.

HPC, Cord Blood has the following inactive ingredients: dimethyl sulfoxide (DMSO), citrate phosphate dextrose (CPD), hydroxyethyl starch, and Dextran 40. When prepared for infusion according to instructions, the infusate contains the following inactive ingredients: Dextran 40, human serum albumin, DMSO, and CPD.

The effectiveness of Bloodworks HPC, Cord Blood, as defined by hematopoietic reconstitution, was demonstrated in one single-arm prospective study and in retrospective reviews of data from an observational database for Bloodworks, and in data in the dockets and public information. Of the 1299 patients in the dockets and public data, 66% (n=862) underwent transplantation as treatment for hematologic malignancy. Results for patients who received a total nucleated cell dose ≥ 2.5 x 107/kg are shown in Table 2. Neutrophil recovery is defined as the time from transplantation to an absolute neutrophil count more than 500 per microliter. Platelet count recovery is the time to a platelet count more than 20,000 per microliter. Erythrocyte recovery is the time to a reticulocyte count greater than 30,000 per microliter. The total nucleated cell dose and degree of HLA match were inversely associated with the time to neutrophil recovery in the docket data.

HPC, Cord Blood is supplied as a cryopreserved cell suspension in a sealed bag containing a minimum of 5 x 108 total nucleated cells with a minimum of 1.25 x 106 viable CD34+ cells in a volume of 25 milliliters (ISBT 128 Product Code S1393, ISBT 128 Facility Identifier Number W1640). The exact pre-cryopreservation nucleated cell content is provided on the container label and accompanying records.

Store HPC, Cord Blood at or below -150°C until ready for thawing and preparation.

Discuss the following with patients receiving HPC, Cord Blood.

• Report immediately any signs and symptoms of acute infusion reactions, such as fever, chills, fatigue, breathing problems, dizziness, nausea, vomiting, headache, or muscle aches.
• Report immediately any signs or symptoms suggestive of graft-versus-host disease, including rash, diarrhea, or yellowing of the eyes.

I. MATERIALS, REAGENTS, AND EQUIPMENT

Materials:

• Large zip-lock plastic bag (sterile interior is recommended)
• Sampling site couplers
• Tubing Clamps
• Sterile 300 mL Transfer Bag (labeled "LMD/25% HSA Solution #1")
• Sterile 300 mL Transfer Bag (labeled "LMD/5% HSA Solution #2")
• Sterile 300 mL Transfer Bag (labeled "Thawed Final Product")
• 14 gauge needles
• 3-5 inch blunt needle (a sterile spinal needle with trochar removed, if available) for emergency product recovery
• Sterile Syringes: 1, 10, 30 and 60 mL
• Sample tubes
• Isopropyl alcohol prep pads, 70% v/v
• Refrigerated gel packs
• Scissors

Reagents:

• 10% Low Molecular Weight Dextran-40 (LMD)
• Human Serum Albumin, USP (HSA), 25% and 5% Solution

Equipment:

• Refrigerator
• Biological Safety Cabinet
• Large water bath, 37°C ± 2°C
• Thermometer
• Di-electric sealer

NOTE: If performing wash step after dilution, then you will also need:

• Sterile Connecting Device
• Centrifuge blood bag adapter and insert (to support low volume bag)
• Centrifuge
• Manual Plasma Extractor
• Scale

Forms:

• Shipping Information
• CBU Summary sheet
• HPC, Cord Blood Release Specifications for Distribution
• HPC, Cord Blood Final Eligibility Determination
• NMDP Receipt of Cord Blood Unit

II. PRODUCT RECEIPT

The HPC, Cord Blood unit is shipped frozen in a dry-shipper that maintains the temperature at ≤ -150°C. The dry shipper is capable of maintaining the temperature of ≤ -150°C for 5 days from the shipping date. If the unit will not be used within that timeframe, transfer the unit into a storage device capable of maintaining the unit at ≤ -150°C.

Temperature during shipment is continuously monitored and a printout can be obtained upon request.

Handle the cryobag with extreme caution when removing it from the liquid nitrogen, metal cassette, and protective overwrap, and during the thaw procedure. Cryobags can be very fragile. This is applicable both during inspection of the product upon arrival and during the thawing process.

1. Upon receiving the shipper, perform the following steps:
   1. Inspect the shipper for tampering or damage prior to opening.
   2. Inspect the temperature monitor on the shipper; mark on the Shipping Information form which light is flashing: Normal, Warning, or Alarm.

      NOTE: If the Warning or Alarm light is flashing, immediately contact Bloodworks. See section VI, Contact Information.

   3. Open the shipper and remove the Tyvek envelope containing a cardboard box.
   4. Open the cardboard box and remove the metal cassette that contains the unit.
   5. Carefully open the cassette. The cassette will contain the unit (cryobag) sealed in an overwrap and padded in an absorbent sheet. Cracks or breakages in the overwrap do not indicate damage to the unit.
   6. Inspect the integrity of the cryobag.

      NOTE: If there is damage to cryobag, replace the cryobag into storage at ≤ -150°C (contain the cryobag in additional overwrap if necessary). Consult with the transplant physician and contact Bloodworks.

   7. Document location of inspection, identity of inspector, and date and time of inspection on the Shipping Information form.
   8. Verify that the unit ID number on the cryobag matches the unit ID number on all applicable paperwork, the unit ID number matches the expected unit for the intended recipient. Document verification on the Shipping Information form.

      NOTE: If there are any discrepancies in unit ID, consult with the transplant physician and contact Bloodworks.

      Store the product in an LN2 Storage vessel that maintains a temperature at or below -150°C.

III. PREPARATION

1. Initial preparation
   1. Coordinate with the clinical team
      1. Confirm the infusion time in advance. Adjust the start time for the thaw so that the unit is available for infusion when the recipient is ready.
      2. Consult with the clinicians about final product volume based on the recipient's weight and possible fluid restrictions, to determine if the Alternate Procedure (see Section IV.4) for DMSO removal and volume reduction should be followed.
   2. Perform clearance of workspace, including disinfection of lab bench and equipment.
   3. Prepare the water bath and verify the temperature is 37°C ± 2°C.
   4. Inspect LMD visually for particulate matter. If crystallization is observed, do not use and discard appropriately.
   5. Pre-cool the LMD, 25% and 5% HSA Solutions in the refrigerator.
   6. Prepare three sterile 300 mL transfer bags.
      1. Using a di-electric sealer, make three seals along the tubing close to the transfer bag, inspect seals for integrity and separate the tubing at the center seal.

         NOTE: If DMSO removal and volume reduction is required, for the 300mL transfer labeled "Thawed Final Product", seal tubing approximately 4-5 inches above the bag to allow for sterile connection in step IV.4.a.

      2. Place the transfer bags into the Biological Safety Cabinet. Insert a sampling coupler in each bag.
2. Prepare Solution #1 with 25% HSA/LMD in the Biological Safety Cabinet.
   1. With a syringe and needle, add 25 mL of cold LMD and 5 mL of cold 25% HSA (per each product being thawed) into a sterile, labeled 300 mL transfer bag.
   2. Mix Solution #1 by inverting and swirling the bag several times thoroughly without creating excessive bubbles.
   3. Store Solution #1 in refrigerator to cool.

      NOTE: The LMD/HSA Solution #1 must be used within 4 hours.

3. Prepare Solution #2 with 5% HSA/LMD in the Biological Safety Cabinet.
   1. With a syringe and needle, add 25 mL of the cold LMD and 25 mL of cold 5% HSA (per each product being thawed) into a sterile, labeled 300 mL transfer bag.
   2. Mix Solution #2 by inverting and swirling the bag several times thoroughly without creating excessive bubbles.
   3. Store Solution #2 in refrigerator to cool.

      NOTE: The LMD/HSA Solution #2 must be used within 4 hours.

IV. PROCEDURE

Reconstitution or simple dilution of HPC, Cord Blood with LMD/HSA solution, using the Thaw and Diluting procedures described below (steps IV.1-IV.3), is recommended. The Alternate Procedure: Washing in Section IV.4 should be followed if the infusion volume and/or DMSO dose (>1 g/kg body weight) are contraindicated for the patient.

1. Thaw HPC, Cord Blood
   1. Verify the identity of the HPC, Cord Blood being thawed.
   2. Using scissors that have been wiped down with alcohol, remove overwrap, detach any segments and store the segments at ≤ -150°C until needed.
   3. Wipe the external surface of the cryobag with isopropyl alcohol. Place the HPC, Cord Blood cryobag inside the large zip-lock plastic bag. Seal the bag closed.

      NOTE: Wiping the external surface of the cryobag with isopropyl alcohol before placing inside the sterile zip-lock bag allows the thawing laboratory to potentially recover the product in the case of an unexpected leak or container failure during thawing.

   4. Completely immerse the bag in the 37°C ± 2°C water bath.
   5. Gently agitate and knead the cryobag until the product reaches a slushy liquid consistency.
   6. Remove bag from the water bath and quickly inspect for tears or leaks.

      NOTE: If container integrity is observed to be compromised, position the cryobag and/or clamp with hemostats to prevent further escape of product. If visible cracks or breaks are observed, consult the transplant physician before proceeding. See Section V for Emergency Product Recovery procedures.

   7. Remove the thawed cryobag from zip-lock bag, wipe with alcohol, and place the cryobag into the Biological Safety Cabinet.
2. Transfer the thawed HPC, Cord Blood into a 300 mL Transfer Bag

   NOTE: Perform the transfer procedure in a Biological Safety Cabinet.

   1. Wipe down the cryobag port covers with alcohol to disinfect.
   2. Cut off port covers with scissors that have been wiped down with alcohol. Only cut through the outer layer of plastic. Do not cut the inner port column.
   3. Insert a sampling site coupler into each port using aseptic technique.
   4. Using a 30 mL syringe with a 14 gauge needle, aseptically withdraw the contents from both compartments into the syringe.
   5. Record the product volume removed (approximately 25 mL).
   6. Using an inserted sampling site coupler, slowly and aseptically transfer the product to a sterile 300 mL transfer bag (labeled as "Thawed Final Product").
   7. Place the product on refrigerated gel packs in the Biological Safety Cabinet.
3. Dilute HPC, Cord Blood

   NOTE: Perform the following dilution procedure in a Biological Safety Cabinet.

   1. Remove the 300 mL Transfer Bag containing Solution #1 from the refrigerator.
   2. Using a 60 mL syringe with a 14 gauge needle, aseptically fill the syringe with Solution #1 of a volume equal to the product volume noted in step 2.e (approximately 25 mL).
   3. Aseptically attach the 60 mL syringe containing Solution #1 to the 300 mL transfer bag containing the thawed product.
   4. Slowly add Solution #1 to the transfer bag while gently agitating the contents of the bag.
   5. Allow product to rest for 5 minutes. Maintain the bag chilled on refrigerated gel packs.
   6. Remove the 300 mL Transfer Bag containing Solution #2 from the refrigerator.
   7. Use a 60 mL syringe with a 14 gauge needle to aseptically fill the syringe with Solution #2, using two times the original product volume noted in step 2.e (approximately 50 mL).
   8. Aseptically attach the 60 mL syringe containing Solution #2 to the 300 mL transfer bag containing the product.
   9. Slowly add Solution #2 to the product while gently agitating the bag. Maintain the syringe and transfer bag chilled on refrigerated gel packs.
   10. Calculate the product volume from the volume noted in steps 2.e, the volume of Solution #1 from step 3.b, and the volume of Solution #2 in step 3.g.
   11. Aseptically remove samples for quality control tests (such as nucleated cell counts, viability testing, CD34 analysis, Colony Forming Units (CFU), and sterility testing) and transfer the samples into appropriately labeled tubes. Document the volume removed.
   12. Subtract the volume removed from the volume noted in step 3.j, thus determining the infusion volume. Document the infusion volume, which will be used for calculating cell numbers.
   13. Immediately transport the product to the clinical infusion site per the facility's SOP.

       NOTE: The recommended expiration time of thawed HPC, Cord Blood product is 4 hours from the time of thaw, if stored at 2-8°C.

4. Alternate Procedure: Washing (DMSO removal and volume reduction)

   Perform all steps of the Thaw and Transfer procedures, and steps 3.a through 3.i of the Dilute procedure as outlined above, and then complete the following steps:

   1. Sterile connect an empty sterile 300 mL transfer bag to the product bag from step 3.i. Place a tubing clip on the tubing close to the product bag.
   2. Place the thawed, diluted product bag and attached 300 mL transfer bag in a blood bag adapter with insert to support the low volume bag and centrifuge at 400 x g for 20 minutes at 4°C.
   3. Place the concentrated HPC, Cord Blood product bag in a manual plasma extractor.
   4. Place the attached empty 300 mL transfer bag on a scale and tare the scale.
   5. Remove the tubing clip. Use the plasma extractor to press off the supernatant into the attached transfer bag until approximately 20 mL cell pellet remains, and clamp the HPC, Cord Blood bag tubing.
   6. Resuspend the cell pellet by gentle agitation. In a Biological Safety Cabinet, use a 30 mL syringe to volumetrically measure the concentrated product. If necessary, add back supernatant to reach a final total volume of 20 mL.
   7. In a Biological Safety Cabinet, aseptically remove samples for quality control tests (such as nucleated cell counts, viability test, CD34 analysis, Colony Forming Units (CFU)) and transfer the samples into appropriately labeled tubes. Do not remove more than 500 microliters. Document the volume removed.
   8. Subtract the sample volume removed in step 4.g from the volume noted in step 4.f, thus determining the infusion volume. Document the infusion volume, which will be used for calculating cell numbers.
   9. In a Biological Safety Cabinet, aseptically add a sampling site coupler to the supernatant transfer bag and remove samples for sterility testing.
   10. Immediately transport the product to the clinical transport site per the facility's SOP.

       NOTE: The recommended expiration time of thawed HPC, Cord Blood product is 4 hours from the time of thaw, if stored at 2-8°C.

V. EMERGENCY PRODUCT RECOVERY IN THE EVENT OF A CONAINER FAILURE

1. Handle the cryobag with extreme caution when removing it from the liquid nitrogen, metal cassette, and protective overwrap and during the thaw proceudre. Cryobags can be very fragile. This is applicable both during inspection of the product upon arrival and during the thawing process.
2. Wipe the external surface of the cryobag with isopropyl alcohol before the cryobag is placed inside a sterile zip-lock bag. This process allows the thawing laboratory to potentially recover the product in the case of an unexpected leak or container failure during thawing or centrifugation.
3. Notify Bloodworks, the transplant physician/team, and local laboratory director immediately if any portion of the product or container seems to be damaged or compromised.
4. If the cryobag is compromised, move further handling into a Biological Safety Cabinet.
5. It is the physician's responsbility to determine whether the product will be used or discarded if the container is compromised at any step of the procedure. If the product is accepted for use, recovery of the product may be attempted as described below.
   1. Prepare an emptry transfer bag with a sampling site coupler.
   2. Use a long (3-5 inch) blunt needle (a sterile spinal needle with trochar removed, if available).
   3. Attach the blunt needle to a sterile 60 mL syringe.
   4. Aspirate the product, replace the blunt needle with a standard needle and inject it into a sterile transfer bag via a sampling site coupler.
   5. Continue processing of the transferred product from the point container failure was identified.
   6. Perform sterility testing on the product sample after final preparation step.

VI. CONTACT INFORMATION

Bloodworks
Cord Blood Services
921 Terry Ave.
Seattle, WA 98104

Hours: 7:30 AM - 6:00 PM, Pacific Standard Time
Phone Number: 206-689-6696
Alternate Phone Number: 1-800-366-2831, ext. 1896

After Hours
Cell Phone Number: 206-231-3108

Distributed by:
Bloodworks
Cord Blood Services
921 Terry Ave.
Seattle, WA 98104
License Number 2042

Principal Display Panel - Carton Label

"W1640 16 999999"

Product: S1393V00

HPC, CORD BLOOD

10% DMSO, Other additives present,
Cryopreserved, Buffy Coat Enriched

See Accompanying Documentation

Total Volume 25.13 mL

Store at -150 C or colder

Expires:

Bloodworks
921 Terry Ave
Seattle, WA 98104

Principal Display Panel - Bag Label

"W1640 16 999999"

Bloodworks
921 Terry Avenue
Seattle, WA 98104

Collection
Date/Time "0160281000"

28 Jan 2016 10:00 PST
(28 Jan, 2016 18:00 UTC)

Do Not Irradiate
Do Not Use Leukoreduction Filters
RX Only

"S1393V00"

HPC, CORD BLOOD

10% DMSO, Other additives present,
Cropreserved, Buffy Coat Enriched

See Accompanying Documentation

Total Volume 25.13 mL

Store at -150 C or colder

US License #: 2042

"6200"

A
Rh Positive

For Intravenous Administration

See package insert for full prescribing
information and instructions for
preparation.

Expiration
Date/Time
"0180281000"

28 Jan 2018 10:00 PST
(28 Jan, 2018 18:00 UTC)

Intended Recipient
Smith, Bob
Recipient ID: 9987-1234-5
Date of Birth: 04 Jul 1970