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Kokkaliaris, K.D. ; Drew, E. ; Endele, M. ; Loeffler, D. ; Hoppe, P.S. ; Hilsenbeck, O. ; Schauberger, B. ; Hinzen, C. ; Skylaki, S. ; Theodorou, M. ; Kieslinger, M. ; Lemischka, I.* ; Moore, K.* ; Schroeder, T.

Identification of factors promoting ex vivo maintenance of mouse hematopoietic stem cells by long-term single-cell quantification.

Blood 128, 1181-1192 (2016)
Publ. Version/Full Text Postprint Supplement DOI PMC
Open Access Green
The maintenance of hematopoietic stem cells (HSCs) during ex vivo culture is an important prerequisite for their therapeutic manipulation. However, despite intense research, culture conditions for robust maintenance of HSCs are still missing. Cultured HSCs are quickly lost, preventing their improved analysis and manipulation. Identification of novel factors supporting HSC ex vivo maintenance is therefore necessary. Co-culture with the AFT024 stroma cell line is capable of maintaining HSCs ex vivo long-term, but the responsible molecular players remain unknown. Here, we use continuous long-term single-cell observation to identify the HSC behavioral signature under supportive or non-supportive stroma co-cultures. We report early HSC survival as a major characteristic of HSC-maintaining conditions. Behavioral screening after manipulation of candidate molecules revealed that the extracellular matrix protein dermatopontin (Dpt) is involved in HSC maintenance. DPT knock-down in supportive stroma impaired HSC survival, while ectopic expression of the Dpt gene or protein in non-supportive conditions restored HSC survival. Supplementing defined stroma- and serum-free culture conditions with recombinant DPT protein improved HSC clonogenicity. These findings illustrate a previously uncharacterized role of Dpt in maintaining HSCs ex vivo.
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Publication type Article: Journal article
Document type Scientific Article
Corresponding Author
Keywords Fibroblast-activation Protein; Self-renewal Capacity; Expansion In-vitro; Bone-marrow Niche; Progenitor Cells; Growth-factor; Adipocyte Differentiation; Stromal Cells; Extracellular-matrix; Targeted Disruption
ISSN (print) / ISBN 0006-4971
e-ISSN 1528-0020
Journal Blood
Quellenangaben Volume: 128, Issue: 9, Pages: 1181-1192 Article Number: , Supplement: ,
Publisher American Society of Hematology
Publishing Place Washington
Non-patent literature Publications
Reviewing status Peer reviewed
Institute(s) Institute of Stem Cell Research (ISF)
Institute of Developmental Genetics (IDG)
Institute of Biological and Medical Imaging (IBMI)