Three-dimensional culture and characterization of mononuclear cells from human bone marrow

Bara, Jennifer J; Herrmann, Marietta; Menzel, Ursula; Benneker, Lorin; Alini, Mauro; Stoddart, Martin J (2015). Three-dimensional culture and characterization of mononuclear cells from human bone marrow. Cytotherapy, 17(4), pp. 458-472. Elsevier 10.1016/j.jcyt.2014.12.011

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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.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Department of Orthopaedic, Plastic and Hand Surgery (DOPH) > Clinic of Orthopaedic Surgery

UniBE Contributor:

Benneker, Lorin Michael

Subjects:

600 Technology > 610 Medicine & health

ISSN:

1465-3249

Publisher:

Elsevier

Language:

English

Submitter:

Fabian Röthlisberger

Date Deposited:

29 Mar 2016 16:28

Last Modified:

01 Apr 2016 16:35

Publisher DOI:

10.1016/j.jcyt.2014.12.011

PubMed ID:

25680302

Uncontrolled Keywords:

FGF(2); bone marrow; bone marrow niche; cell transplantation; mesenchymal stem cells; mononuclear cells

BORIS DOI:

10.7892/boris.77287

URI:

https://boris.unibe.ch/id/eprint/77287

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