Human adipose-tissue derived stromal cells in combination with hypoxia effectively support ex vivo expansion of cord blood haematopoietic progenitors.
The optimisation of haematopoietic stem and progenitor cell expansion is on demand in modern cell therapy. In this work, haematopoietic stem/progenitor cells (HSPCs) have been selected from unmanipulated cord blood mononuclear cells (cbMNCs) due to adhesion to human adipose-tissue derived stromal cells (ASCs) under standard (20%) and tissue-related (5%) oxygen. ASCs efficiently maintained viability and supported further HSPC expansion at 20% and 5% O2. During co-culture with ASCs, a new floating population of differently committed HSPCs (HSPCs-1) grew. This suspension was enriched with СD34+ cells up to 6 (20% O2) and 8 (5% O2) times. Functional analysis of HSPCs-1 revealed cobble-stone area forming cells (CAFCs) and lineage-restricted colony-forming cells (CFCs). The number of CFCs was 1.6 times higher at tissue-related O2, than in standard cultivation (20% O2). This increase was related to a rise in the number of multipotent precursors - BFU-E, CFU-GEMM and CFU-GM. These changes were at least partly ensured by the increased concentration of MCP-1 and IL-8 at 5% O2. In summary, our data demonstrated that human ASCs enables the selection of functionally active HSPCs from unfractionated cbMNCs, the further expansion of which without exogenous cytokines provides enrichment with CD34+ cells. ASCs efficiently support the viability and proliferation of cord blood haematopoietic progenitors of different commitment at standard and tissue-related O2 levels at the expense of direct and paracrine cell-to-cell interactions.
Institute of Biomedical Problems, Russian Academy of Sciences, Moscow, Russia.
Elena R
ER
Andreeva
Institute of Biomedical Problems, Russian Academy of Sciences, Moscow, Russia; Institute of Experimental Cardiology, Cardiology Research Center, Moscow, Russia.
Irina V
IV
Andrianova
Institute of Biomedical Problems, Russian Academy of Sciences, Moscow, Russia.
Elena V
EV
Sotnezova
Faculty of Basic Medicine, Moscow State University, Moscow, Russia.
Sergey V
SV
Buravkov
Institute of Biomedical Problems, Russian Academy of Sciences, Moscow, Russia.
Polina I
PI
Bobyleva
Institute of Experimental Cardiology, Cardiology Research Center, Moscow, Russia.
Yury A
YA
Romanov
Institute of Biomedical Problems, Russian Academy of Sciences, Moscow, Russia.
Ludmila B
LB
Buravkova
D000273:Adipose Tissue / Q000166:cytology; D002448:Cell Adhesion; D015687:Cell Hypoxia; D049109:Cell Proliferation; D018920:Coculture Techniques; D005312:Fetal Blood / Q000166:cytology*; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism*
D010100:Oxygen