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ctt-journal > Aizenstadt A.1 et al. (Abstract)

Aizenstadt A.1 et al. (Abstract)

Cellular Therapy and Transplantation (CTT), Vol. 3, No. 12
doi: 10.3205/ctt-2011-No12-abstract80

© 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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Preparation of cord blood mesenchymal stem cells for transplantation: expansion and quality control

Alexandra A. Aizenstadt, Irina L. Trofimova, 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

Mesenchymal stem cells (MSCs) offer well-known immunosuppressive properties and can be successfully used during devastating autoimmune disease treatment, such as graft-versus-host disease. Cord blood is known to be one of the most rich sources of MSCs. However the amount of MSCs in CB never exceeds 0.0005% of all nucleated cells.

The development of the most effective CB MSCs expansion method, and the establishment of quality control procedures were the aims of this investigation. 

We selected optimal conditions for CB MSCs expansion that provided their maximal viability and proliferative activity. MSCs were cultured on AdvanceStem Media for no longer than 4 passages. The first passage was on 24–28 day after explantation. Then the cell culture was seeded every 5–7 days, with an initial density of 1.3x103 cells/sm2. In order to perform quality control on the cells, cytogenetic assay and immunophenotyping were applied at every passage for one part of the MSCs cell culture. After each passage the conditional media was tested for sterility and the presence of Mycoplasma sp. For analysis of immunosuppressive internals CB MSCs cultures were cultivated together with stimulated lymphocytes of peripheral blood from intact donors that were marked by CFSE.

Clear MSCs populations were obtained; their phenotype didn’t change during cultivation and is described as CD34-, CD45-, CD44+, CD90+, and CD105+. No alterations in chromosome amount and/or structure were detected in any sample during karyotyping. No alteration in chromosome structure or amount was detected in samples. Neither bacterial nor fungal contamination was detected in samples. CB MSCs suppressed lymphocyte proliferation, i.e., possessed prominent immunosuppressive properties.

Keywords: cord blood mesenchymal stem cells, expansion, quality control