The Effect of Alterations of Hemostatic Proteins on the Yield of Progenitor Cells in Anticoagulated and Elapsed Cord Blood

Park, Hyun Woo; Lee, Young Ho; Lee, Young Ah; Kim, Kyeong Hee; Han, Jin Yeong

Abstract

PURPOSE: We assessed the changes of the activities of hemostatic variables and also their relationship with the yield of progenitor cells by analysing the effect of urokinase on the elapsed cord blood (CB). METHPDS: We collected 34 CBs using syringes anticoagulated with acid-citrate-dextrose (ACD). The 19 CBs which were held evenly at ambient temperature and 4oC, were separated immediately, 24, 48 and 72 hours after collection, and then the total nucleated cell (TNC) and CD34+ cell counts were measured. We had frozen plasma from every samples and analysed the activities of antithrombin (AT) III, protein C, plasminogen and alpha2 antiplasmin using an MLA (Medical Laboratory Automation) coagulation analyzer. For the other 15 CBs, we assessed the effect of urokinase by comparing TNC and CD34+ cell counts between fresh and 48 hour-elapsed CB treated with or without 10,000 IU/mL of urokinase.
RESULTS
The significant loss of TNC and CD34+ cells continued to 24 hours after collection in CB stored at 4degrees C, but loss of TNC began only after 24 hours at ambient temperature. The activity of protein C was decreased significantly until 24 hours after collection, particularly in CB stored at 4degrees C than at ambient temperature. The activity of alpha2 antiplasmin was decreased until 24 hours in CB stored at 4degrees C and from 24 hours in CB stored at ambient temperature. TNC and CD34+ cell counts after separation in 48 hour-elapsed CB were significantly higher in urokinase-treated samples than untreated samples.
CONCLUSION
The alterations of the activities of hemostatic proteins could be an important factor of coagulability, particularly even in anticoagulated, which also affect the yield of progenitor cells in processed CB.