Collection strategies and cryopreservation of umbilical cord blood

Abstract

The aim of this study was to compare (a) two different umbilical cord blood (UCB) collection methods while the placenta is still in the uterus (in utero), and (b) to evaluate the efficacy of four cryopreservation protocols based on UCB haematopoiestic stem cell (HSC) recovery. We analysed UCB samples collected with our original collection system designed for active Syringe/Flush/Syringe method or by standard in utero method. For comparing different cryopreservation procedures, dimethyl sulphoxide (DMSO) at final concentration of 5 and 10% was used and combined with our own controlled-rate or uncontrolled-rate cryopreservation. A total of 99 samples were collected. A significantly higher UCB volume, total nucleated cell and mononuclear cell were seen following the first collection strategy (n= 49; mean +/- SD, 103 +/- 35.4 mL; 12.34 +/- 5.27 x 10(8); 595 +/- 3.47 x 10(6)) vs. the second strategy (n= 50; 86 +/- 29.3 mL; 9.87 +/- 4.47; 424 +/- 2.82 x 10(6)) respectively (P < 0.01). The discard rate was 14% for the first and 36% for the second collection strategy (P < 0.01). It was shown that the most efficient procedure was the controlled-rate protocol combined with lower (5%) DMSO concentration. Using active Syringe/Flush/Syringe method, we collected UCB with greater volumes and with lower discard rate compared to the standard by gravity technique. The data presented also showed much better recovery of UCB cells when controlled-rate freezing procedure and 5% DMSO were combined.

Concise Review: Isolation and Characterization of Cells from Human Term Placenta: Outcome of the First International Workshop on Placenta Derived Stem Cells

“…Different methods for the reduction of red blood cells, plasma volume, and cryopreservation exist [111]. Cord blood products containing cryoprotectants (e.g., dimethyl sulfoxide [DMSO]) are frozen at a controlled-rate and can be stored in liquid nitrogen for at least 15 years without the loss of their engraftment potential in vivo [110,112].…”

Incorporation of Placenta Extract Into Liposomal Drug Delivery System

“…[42] Cord blood products containing cryoprotectants (e.g., dimethyl sulfoxide [DMSO]) are frozen at a controlled-rate and can be stored in liquid nitrogen for at least 15 years without the loss of their engraftment potential in vivo. [43] www.wjpr.net The use of amniotic membrane has history spanning almost 100 years. The first reported clinical use of amniotic membrane was in 1910, when it was applied in skin transplantation.…”

Cord Blood Stem Cell Cryopreservation

“…Because UCB HPCs were thought to be very similar to their bone marrow-derived counterparts, initial protocols were derived from those used for the preservation of bone marrow and peripheral blood [19]. However, there is not substantial evidence that such protocols are optimal for UCB sources [20]. To develop a successful cell cryopreservation strategy, the key events inherent to freezing a biological cell must be considered, and each potential source of damage should be mitigated if at all possible.…”