The effects of co-culture duration of cord blood cells with adipose tissue-derived stromal cells on hematopoietic precursors> amplification

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Abstract

Umbilical cord blood is considered as a valuable source of hematopoietic stem and progenitor cells (CB-HSPCs). The number of latter may be significantly enriched with ex vivo expansion. Thus, the optimization of culture conditions is essential for in vitro manipulations. Recently we have demonstrated that CB-HSPCs may be separated from unmanipulated CB nucleated cells through the adhesion to adipose tissue-derived MSCs. Further coculture was resulted in raizing of new polulation of floating CB-HSPCs significantly ehriched in primitive progentors The goal of this study was to optimize above mentioned protocol To determine the optimal conditions for adhesion and multiplication of CB-HSPCs, nucleated CB cells were co-cultured on adipose-tissue MSC layer for short (1-3 hours) and long-term (24-72 hours) duration Unattached cells were removed, adherent CB-HSPCs were further cultured for 72 hours, resulted in formation of floating population of CB-HSPCs. in each time point the number of attached CB-HSPCs, newly formed floating CB-HSPCs, CD34+ cells and CFUs among latter was examined. After 72 hours of nucleated CB cells co-culture, the number of adherent CD34+ cells peaked and was over than 70% of total CD34+ cells among nucleated CB cell samples Proposed experimental design has provided 4-fold enrichment of primitive CD34+ and 6-fold of CFUs number among newly formed HSPCs. BFU-Es comprised 80-90% of total CFUs regardless of time of nucleated CB cells coculture. Thus, 3 days of nucleated CB cells/adipose tissue-derived mesenchymal stromal cells co-culture provided peak of CD34+ cells' adhesion, amplification of latter resulted in rising of population maximally enriched both with undifferentiated and committed hematopoietic precursors

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About the authors

E. R Andreeva

Institute of Biomedical Problems, Russian Academy of Sciences

Email: andreeva_er@mail.ru

I. V Andrianova

Institute of Biomedical Problems, Russian Academy of Sciences; Institute of Experimental Cardiology, Cardiology Research Center

A. N Gornostaeva

Institute of Biomedical Problems, Russian Academy of Sciences

P. I Bobyleva

Institute of Biomedical Problems, Russian Academy of Sciences

E. E Balashova

Institute of Experimental Cardiology, Cardiology Research Center; Cord Blood Bank “CryoCenter,"

L. B Buravkova

Institute of Biomedical Problems, Russian Academy of Sciences

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