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Three-Dimensional Perfusion Culture of Human Bone Marrow Cells and Generation of Osteoinductive Grafts

^a Departments of Surgery and of Research, University Hospital Basel, Basel, Switzerland;
^b Department of Oncology, Biology and Genetics, University of Genova, Genova, Italy

Key Words. Bone marrow stromal cells • Bone marrow cells • Colony formation • Expansion • Hematopoiesis • Mesenchymal stem cells • Osteoprogenitor • Tissue regeneration

Correspondence: Ivan Martin, Ph.D., Institute for Surgical Research and Hospital Management, University Hospital Basel, Hebelstrasse 20, ZLF, Room 405, 4031 Basel, Switzerland. Telephone: 41-61-265-2384; Fax: 41-61-265-3990; e-mail: imartin{at}uhbs.ch

Three-dimensional (3D) culture systems are critical to investigate cell physiology and to engineer tissue grafts. In this study, we describe a simple yet innovative bioreactor-based approach to seed, expand, and differentiate bone marrow stromal cells (BMSCs) directly in a 3D environment, bypassing the conventional process of monolayer (two-dimensional [2D]) expansion. The system, based on the perfusion of bone marrow–nucleated cells through porous 3D scaffolds, supported the formation of stromal-like tissues, where BMSCs could be cocultured with hematopoietic progenitor cells in proportions dependent on the specific medium supplements. The resulting engineered constructs, when implanted ectopically in nude mice, generated bone tissue more reproducibly, uniformly, and extensively than scaffolds loaded with 2D-expanded BMSCs. The developed system may thus be used as a 3D in vitro model of bone marrow to study interactions between BMSCs and hematopoietic cells as well as to streamline manufacture of osteoinductive grafts in the context of regenerative medicine.

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