Secreted factors from mouse embryonic fibroblasts maintain repopulating function of single cultured hematopoietic stem cells
Sandra Romero Marquez,
Franziska Hettler,
Renate Hausinger,
Christina Schreck,
Theresa Landspersky,
Lynette Henkel,
Corinne Angerpointner,
Ihsan E. Demir,
Matthias Schiemann,
Florian Bassermann,
Katharina S. Götze,
Rouzanna Istvánffy,
Robert A.J. Oostendorp
Affiliations
Sandra Romero Marquez
Technical University of Munich, Klinikum rechts der Isar, Clinic and Polyclinic for Internal Medicine III, Munich, Germany; Technical University of Munich, Klinikum rechts der Isar, Department of Surgery, Munich
Franziska Hettler
Technical University of Munich, Klinikum rechts der Isar, Clinic and Polyclinic for Internal Medicine III, Munich
Renate Hausinger
Technical University of Munich, Klinikum rechts der Isar, Clinic and Polyclinic for Internal Medicine III, Munich
Christina Schreck
Technical University of Munich, Klinikum rechts der Isar, Clinic and Polyclinic for Internal Medicine III, Munich
Theresa Landspersky
Technical University of Munich, Klinikum rechts der Isar, Clinic and Polyclinic for Internal Medicine III, Munich
Lynette Henkel
Technical University of Munich, Flow Cytometry Unit of the Technical University Munich, Institute for Medical Microbiology, Immunology and Hygiene (CyTUM-MIH), Munich
Corinne Angerpointner
Technical University of Munich, Flow Cytometry Unit of the Technical University Munich, Institute for Medical Microbiology, Immunology and Hygiene (CyTUM-MIH), Munich
Ihsan E. Demir
Technical University of Munich, Klinikum rechts der Isar, Department of Surgery, Munich
Matthias Schiemann
Technical University of Munich, Flow Cytometry Unit of the Technical University Munich, Institute for Medical Microbiology, Immunology and Hygiene (CyTUM-MIH), Munich
Florian Bassermann
Technical University of Munich, Klinikum rechts der Isar, Clinic and Polyclinic for Internal Medicine III, Munich, Germany; German Cancer Consortium (DKTK), Heidelberg
Katharina S. Götze
Technical University of Munich, Klinikum rechts der Isar, Clinic and Polyclinic for Internal Medicine III, Munich, Germany; German Cancer Consortium (DKTK), Heidelberg
Rouzanna Istvánffy
Technical University of Munich, Klinikum rechts der Isar, Clinic and Polyclinic for Internal Medicine III, Munich, Germany; Technical University of Munich, Klinikum rechts der Isar, Department of Surgery, Munich
Robert A.J. Oostendorp
Technical University of Munich, Klinikum rechts der Isar, Clinic and Polyclinic for Internal Medicine III, Munich
Hematopoietic stem cell self-renewal, proliferation, and differentiation are independently regulated by intrinsic as well as extrinsic mechanisms. We previously demonstrated that murine proliferation of hematopoietic stem cells is supported in serum-free medium supplemented with two growth factors, stem cell factor and interleukin 11. The survival of hematopoietic stem cells is additionally improved by supplementing this medium with two more growth factors, neural growth factor and collagen 1 (four growth factors) or serum-free medium conditioned by the hematopoietic stem cell-supportive stromal UG26-1B6 cells1. Here, we describe a robust and versatile alternative source of conditioned medium from mouse embryonic fibroblasts. We found that this conditioned medium supports survival and phenotypical identity of hematopoietic stem cells, as well as cell cycle entry in single cell cultures of CD34- CD48- CD150+ Lineage- SCA1+ KIT+ cells supplemented with two growth factors. Strikingly, in comparison with cultures in serum-free medium with four growth factors, conditioned medium from mouse embryonic fibroblasts increases the numbers of proliferating clones and the number of Lineage- SCA1+ KIT+ cells, both with two and four growth factors. In addition, conditioned medium from mouse embryonic fibroblasts supports self-renewal in culture of cells with short- and long-term hematopoiesis-repopulating ability in vivo. These findings identify conditioned medium from mouse embryonic fibroblasts as a robust alternative serumfree source of factors to maintain self-renewal of in vivo-repopulating hematopoetic stem cells in culture.