Jennifer J Bara1, Marietta Herrmann1, Ursula Menzel1, Lorin Benneker2, Mauro Alini1, Martin J Stoddart3. 1. AO Research Institute Davos, Davos Platz, Switzerland. 2. Department of Orthopaedic Surgery, Inselspital, Bern University Hospital and University of Bern, Bern, Switzerland. 3. AO Research Institute Davos, Davos Platz, Switzerland. Electronic address: martin.stoddart@aofoundation.org.
Abstract
BACKGROUND AIMS: The diverse phenotypic changes and clinical and economic disadvantages associated with the monolayer expansion of bone marrow-derived mesenchymal stromal cells (MSCs) have focused attention on the development of one-step intraoperative cells therapies and homing strategies. The mononuclear cell fraction of bone marrow, inclusive of discrete stem cell populations, is not well characterized, and we currently lack suitable cell culture systems in which to culture and investigate the behavior of these cells. METHODS: Human bone marrow-derived mononuclear cells were cultured within fibrin for 2 weeks with or without fibroblast growth factor-2 supplementation. DNA content and cell viability of enzymatically retrieved cells were determined at days 7 and 14. Cell surface marker profiling and cell cycle analysis were performed by means of multi-color flow cytometry and a 5-ethynyl-2'-deoxyuridine incorporation assay, respectively. RESULTS: Total mononuclear cell fractions, isolated from whole human bone marrow, was successfully cultured in fibrin gels for up to 14 days under static conditions. Discrete niche cell populations including MSCs, pericytes and hematopoietic stem cells were maintained in relative quiescence for 7 days in proportions similar to that in freshly isolated cells. Colony-forming unit efficiency of enzymatically retrieved MSCs was significantly higher at day 14 compared to day 0; and in accordance with previously published works, it was fibroblast growth factor-2-dependant. CONCLUSIONS: Fibrin gels provide a simple, novel system in which to culture and study the complete fraction of bone marrow-derived mononuclear cells and may support the development of improved bone marrow cell-based therapies.
BACKGROUND AIMS: The diverse phenotypic changes and clinical and economic disadvantages associated with the monolayer expansion of bone marrow-derived mesenchymal stromal cells (MSCs) have focused attention on the development of one-step intraoperative cells therapies and homing strategies. The mononuclear cell fraction of bone marrow, inclusive of discrete stem cell populations, is not well characterized, and we currently lack suitable cell culture systems in which to culture and investigate the behavior of these cells. METHODS:Human bone marrow-derived mononuclear cells were cultured within fibrin for 2 weeks with or without fibroblast growth factor-2 supplementation. DNA content and cell viability of enzymatically retrieved cells were determined at days 7 and 14. Cell surface marker profiling and cell cycle analysis were performed by means of multi-color flow cytometry and a 5-ethynyl-2'-deoxyuridine incorporation assay, respectively. RESULTS: Total mononuclear cell fractions, isolated from whole human bone marrow, was successfully cultured in fibrin gels for up to 14 days under static conditions. Discrete niche cell populations including MSCs, pericytes and hematopoietic stem cells were maintained in relative quiescence for 7 days in proportions similar to that in freshly isolated cells. Colony-forming unit efficiency of enzymatically retrieved MSCs was significantly higher at day 14 compared to day 0; and in accordance with previously published works, it was fibroblast growth factor-2-dependant. CONCLUSIONS: Fibrin gels provide a simple, novel system in which to culture and study the complete fraction of bone marrow-derived mononuclear cells and may support the development of improved bone marrow cell-based therapies.
Authors: Caroline Moser; Katie Bardsley; Alicia J El Haj; Mauro Alini; Martin J Stoddart; Jennifer J Bara Journal: Front Bioeng Biotechnol Date: 2018-11-15
Authors: Sebastian Wangler; Amir Kamali; Christina Wapp; Karin Wuertz-Kozak; Sonja Häckel; Claudia Fortes; Lorin M Benneker; Lisbet Haglund; R Geoff Richards; Mauro Alini; Marianna Peroglio; Sibylle Grad Journal: Stem Cell Res Ther Date: 2021-01-07 Impact factor: 6.832
Authors: Ander Abarrategi; Syed A Mian; Diana Passaro; Kevin Rouault-Pierre; William Grey; Dominique Bonnet Journal: J Exp Med Date: 2018-02-16 Impact factor: 14.307