HEMACORD consists of hematopoietic progenitor cells, monocytes, lymphocytes, and granulocytes derived from umbilical cord blood of donors and is intended for intravenous infusion. Blood recovered from umbilical cord and placenta is volume-reduced and partially depleted of red blood cells and plasma.
Blood remaining in the delivered placenta and umbilical cord is retrieved
by inserting a 16-gauge needle of a standard 450-ml blood donor set containing CPD A anticoagulant (citrate/phosphate/dextrose/adenine), into the umbilical vein of the delivered placenta and letting it drain by gravity into the blood bag, usually retrieving <170 ml.
Hydroxyethyl starch (HES, 6% wt/vol) is added to a final concentration of 1.2%. A leukocyte-rich supernatant is then separated by centrifuging the mixture in the original collection blood bag (50 x g for 5 min at 10°C). The leukocyte-rich supernatant is expressed from the bag into a 150-ml Plasma Transfer bag and centrifuged (400 x g for 10 min) to sediment the cells. Surplus supernatant plasma is transferred into a second Plasma Transfer bag without severing the connecting tube.
Finally, the sedimented leukocytes are resuspended in supernatant plasma to a total volume of 20 ml.
Dimethyl sulfoxide (DMSO) is used at a final concentration of 10% (vol/vol). The required volume of sterile, chilled DMSO solution is added to the blood bag, over the course of 15 min, by using a syringe pump and an orbital mixer to assure smooth but vigorous mixing kept cold with wet ice throughout the addition. When the concentration of DMSO reaches 10%, cell suspensions are transferred to Cryocyte freezing containers placed in aluminum canisters, and then deposited horizontally on a level surface inside a -80°C freezer. When their temperatures are below -50°C, the units are transferred to the liquid nitrogen freezer for storage.
EMBRYONIC DEVELOPMENT & STEM CELL COMPENDIUM
