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Evaluation of committed and primitive cord blood progenitors after expansion on adipose stromal cells

Andreeva, E.R., Andrianova, I.V., Gornostaeva, A.N., Gogiya, B.Sh., Buravkova, L.B.
Cell and tissue research 2018 v.372 no.3 pp. 523-533
blood, coculture, hematopoiesis, hematopoietic stem cells, humans, phenotype, stromal cells, therapeutics, umbilical cord
Umbilical cord blood mononuclear fraction is a valuable source of hematopoietic stem and progenitor cells (CB HSPCs). The rarity of this population is a serious limitation of its application in cell therapy. Ex vivo expansion enables to significantly amplify the number of hematopoietic precursors of different commitment. Here, we expand CB MNCs in co-culture with human adipose tissue-derived stromal cells (ASCs) to enrich HSPCs and describe phenotypic features of newly formed hematopoietic populations. The CD34⁺-HSPCs demonstrated 6-fold enrichment with 9000 CFUs per 50 × 10³ HSPCs on average. A part of the floating HSPCs were bearing lineage markers, while others were primitive precursors (CD133⁻/CD34⁺). Among ASC-associated HSPCs, two subsets of cord blood-borne cells were revealed: СD90⁺/СD45⁻ and СD90⁺/СD45⁺. The proportion of CD3⁺/CD8⁺ and NK-T as well as CD25⁺ and HLA-DR⁺ Т cells among СD90⁺/СD45⁻ cells was significantly higher compared to MNCs and floating HSPCs. More than 80% of CD45⁺/СD90⁺ HSPCs were identified as late primitive precursors (CD133⁻/CD34⁺). Thus, CB MNC expansion in the presence of ASCs provides the generation of both lineage committed lymphoid progenitors and CD34⁺/CD133⁻ primitive HSPCs. Substantially enriched with primitive precursors, ASC-associated HSPCs could be considered as a perspective tool for a long-term restoration of hematopoiesis in various hematologic disorders.