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Barkhatov et al. abstract

Please cite this article as follows: Barkhatov IM, Roumiantsev SA, Vladimirskaya EB, Afanasyev BV. Composition and functional properties of monolayer cell culture from human umbilical cord blood. Cell Ther Transplant. 2008;1:e.000026.01. doi:10.3205/ctt-2008-en-000026.01

 

Composition and functional properties of monolayer cell culture from human umbilical cord blood

Barkhatov I.M., Roumiantsev S.A., Vladimirskaya E.B., Afanasyev B.V.
Abstract

Objectives: It’s known that during cultivation, adherent cells of umbilical cord blood (UCB) form a monolayer reminiscent, in its composition, of the stromal monolayer of bone marrow (BM) culture. However, the presence of mesenchymal stem cells (MSCs) in UCB still remains uncertain. This study was performed to investigate the composition and some functional characteristics of MSC-like cell populations revealed in the cord blood monolayer culture.
Materials and methods: Forty-three human UCB samples were under study. All the samples were obtained during full-term deliveries. To produce monolayer cultures, mononuclear cell fractions from UCB were cultivated in a culture medium containing DMEM with 20% FCS, supplied with 1% Pen/Strep. Phenotypic patterns of UCB culture were assessed with a panel of monoclonal antibodies specific for CD34; CD117; CD45; CD14; CD3; CD19; CD31; CD90; HLA DR; and HLA ABC. To determine the functional characteristics of MSCs derived from UCB culture, their differentiation ability and stimulation of hematopoietic colony formation activity were evaluated.
Results: In most cases, the cultures of plastic-adherent cells proved to be heterogeneous. Both spindle-shaped and polygonal cells were observed. In some samples, clonal growth could be detected. However, the number of fibroblastoid cells did not increase 100 cells per colony. Large colonies were registered in three UCB samples of the 43 under study. As evidenced by immune phenotyping, the monolayer UCB cultures were rather polymorphic and dissimilar in each sample. Most of the cells present in the cultures were macrophages (CD45+). However, we also found different amounts of presumably mesenchymal cells, including cells with an endothelial phenotype (CD34+CD31+).
Specific staining showed that the cells from a UCB monolayer culture have the capacity to differentiate into adipocytes and osteoblasts. In some cultures, however, induction of differentiation lead to the detachment of a major cell fraction. Hemostimulatory ability of UCB monolayer cultures depended on the phenotype composition of the monolayer culture. CD45+ and CD14+ cells, evidently, are stimulatory for granulocyte-macrophage colony formation. Moreover, levels of non-hematopoietic subpopulations (CD90+CD31-) in UCB cultures showed a direct correlation with the numbers of CFU-GM colonies produced.
Conclusion: UCB contains a subpopulation of non-hematopoietic cells possessing phenotypic and some functional characteristics of bone marrow derived mesenchymal stem cells. However, the low content and variable numbers of such cells provide some doubts on the viability of UCB as an alternative source for MSC.

Keywords: umbilical cord blood, mesenchymal stem cells (MSC), immunophenotyping, cell culture

Please cite this article as follows: Barkhatov IM, Roumiantsev SA, Vladimirskaya EB, Afanasyev BV. Composition and functional properties of monolayer cell culture from human umbilical cord blood. Cell Ther Transplant. 2008;1:e.000026.01. doi:10.3205/ctt-2008-en-000026.01


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