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ctt-journal > Korovina et al. (Abstract)

Korovina et al. (Abstract)

Cellular Therapy and Transplantation (CTT), Vol. 3, No. 12
doi: 10.3205/ctt-2011-No12-abstract85

© The Authors. This abstract is provided under the following license: Creative Commons Attribution 3.0 Unported

Abstract accepted for "5th Raisa Gorbacheva Memorial Meeting Hematopoietic Stem Cell Transplantation in Children and Adults", Saint Petersburg, Russia, September 18–20, 2011

Preliminary Program

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The selection of an optimal method of umbilical cord blood sample preparation for transplantation

Kseniya V. Korovina, Alexandrina S. Khrupina, Kseniya V. Shunkina, Alexander B. Smolyaninov

Research laboratory of cell technologies of the North-Western State Medical University named after I.I. Mechnikov, St. Petersburg, Russia; "Stem Cell Bank Pokrovsky", St. Petersburg, Russia

Correspondence:Alexander B. Smolyaninov, "Stem Cell Bank Pokrovsky", Bolshoi Prospect Vasilevski Ostrov 85, St. Petersburg, 199106, Russia, E-mail: stemcellbank@spam is badinbox.ru

Abstract

Purpose: The purpose of this study was to determine the optimal method for cord blood sample preparation for transplantation in certain conditions in hematology.

Materials and methods: Seventy-seven cord blood samples collected during delivery and before the delivery of the placenta were processed by two methods: double centrifugation (using Rotanta 460RS, Hettich, Germany) (n=38) (group M) and automatic separation (using Sepax S-100, Biosystem, Switzerland) (n=39) (group S). After processing, the samples were stored in liquid nitrogen at -196°C. Once removed from storage the samples were divided into three subgroups, depending on the thawing method: dilution and manual washing from DMSO (subgroup A), automatic thawing and washing using Sepax S-100 (Biosystem, Switzerland) (subgroup B), and without washing (subgroup C). The number of nucleated cells per milliliter (TNC/ml) was the parameter for efficient separation and thawing. The measurements were performed using the Beckman Coulter FC500 flow cytometer (Beckman Coulter, USA). The cell yield was calculated by the number of TNC/ml before cryopreservation.

Results: After thawing, the yield of cells in group M was: 25.7±7,88 TNC/ml (74%), 39.6±2.44 TNC/ml (85.99%), and 45,7±0.44 TNC/ml (92.78%) in subgroups A, B, and C respectively. The yield of cells in group S was 29.48±10.26 TNC/ml (82.47%), 41.57±11.10 TNC/ml (88%), and 48.95±25.63 TNC/ml (98.76%) in subgroups A, B, and C respectively.

Conclusions: The data obtained shows that the best yield of cells after thawing is provided by the absence of washing. If the decision is made to wash the cord blood sample for the prevention of adverse toxic effects of DMSO, the use of an automatic separator will be the most effective.

Keywords: umbilical cord blood, thawing, separation, nucleated cells, cryoprotectant