An efficient method for the cryopreservation of fetal human liver hematopoeitic progenitor cells

Abstract

The use of human hematopoietic progenitor cells (HPC) for transplantation requires efficient recovery methods and cryopreservation procedures. The purpose of this study was to determine cryopreservation techniques for fetal human liver (FHL) CD34(+) cells. We assessed FHL HPC recovery efficiency after freezing and thawing by viability testing, fluorescence-activated cell sorting analysis, and colony-forming ability under different conditions. We also determined optimal cell freezing concentrations and the effect of rate-controlled freezing on cell recovery. Lastly, cell recovery after varying freezing time periods was examined. Our results indicated that optimal cell recovery occurs when: A) cryopreservation medium consists of either 5% dimethylsulphoxide (DMSO) or 10% DMSO in combination with either 20% fetal bovine serum (FBS) or 70% FBS and when Iscove's modified Dulbecco's medium consists of not more than 10% DMSO; B) a rate-controlled freezing device container is used; C) CD34(+) cells are frozen at a concentration of 1 x 10(6)/ml, and D) a thawing temperature of 37 degrees C is used. These observations indicate that cryopreservation of FHL HPC is possible for up to 18 months in optimal conditions without losing hematopoietic activity.