Prolonged ex vivo culture of human bone marrow mesenchymal stem cells influences their supportive activity toward NOD/SCID-repopulating cells and committed progenitor cells of B lymphoid and myeloid lineages

• Published in: Haematologica (Roma) Vol. 95; no. 1; pp. 47 - 56
• Main Authors: BRIQUET, Alexandra, DUBOIS, Sophie, BEKAERT, Sandrine, DOLHET, Marie, BEGUIN, Yves, GOTHOT, André
• Format: Journal Article Web Resource
• Language: English
• Published: Pavia Ferrata Storti Foundation 01.01.2010
• Subjects: Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy; Animals; Biological and medical sciences; Bone Marrow Cells - cytology; Bone Marrow Cells - physiology; Bone marrow, stem cells transplantation. Graft versus host reaction; Cell Culture Techniques - methods; Cell Differentiation - physiology; Cell Lineage - physiology; Cell Proliferation; Hematologic and hematopoietic diseases; Hematology; human bone marrow mesenchymal stem cells; Human health sciences; Humans; Hématologie; Lymphoid Progenitor Cells - cytology; Lymphoid Progenitor Cells - physiology; Medical sciences; Mesenchymal Stromal Cells - cytology; Mesenchymal Stromal Cells - physiology; Mice; Mice, Inbred NOD; Mice, SCID; Myeloid Progenitor Cells - cytology; Myeloid Progenitor Cells - physiology; NOD/SCID-repopulating cells; Original; progenitor cells of B lymphoid; Sciences de la santé humaine; Time Factors; Transfusions. Complications. Transfusion reactions. Cell and gene therapy; Human; Cell proliferation; Cell culture; Lymphoid cell; Hematology; Stem cell; Hematopoietic cell; NOD/SCID mouse; B-Lymphocyte; Mesenchymal cell; Myeloid cell; progenitor cells of B lymphoid; NOD/SCID-repopulating cells; Ex vivo; Bone marrow; human bone marrow mesenchymal stem cells; Prolonged
• DOI: 10.3324/haematol.2009.008524
• ISSN: 0390-6078; 1592-8721

Bone marrow mesenchymal stem cells support proliferation and differentiation of hematopoietic progenitor cells in vitro. Since these cells constitute a rare subset of bone marrow cells, mesenchymal stem cell preparations for clinical purposes require a preparative step of ex vivo multiplication. The aim of our study was to analyze the influence of culture duration on mesenchymal stem cell supportive activity. Mesenchymal stem cells were expanded for up to ten passages. These cells and CD34+ cells were seeded in cytokine-free co-cultures after which the phenotype, clonogenic capacity and in vivo repopulating activity of harvested hematopoietic cells were assessed. Early passage mesenchymal stem cells supported hematopoietic progenitor cell expansion and differentiation toward both B lymphoid and myeloid lineages. Late passage mesenchymal stem cells did not support hematopoietic progenitor cell and myeloid cell outgrowth but maintained B-cell supportive ability. In vitro maintenance of NOD/SCID mouse repopulating cells cultured for 1 week in contact with mesenchymal stem cells was effective until the fourth passage of the mesenchymal cells and declined thereafter. The levels of engraftment of CD34(+) cells in NOD/SCID mice was higher when these cells were co-injected with early passage mesenchymal stem cells; however mesenchymal cells expanded beyond nine passages were ineffective in promoting CD34(+) cell engraftment. Non-contact cultures indicated that mesenchymal stem cell supportive activity involved diffusible factors. Among these, interleukins 6 and 8 contributed to the supportive activity of early passage mesenchymal stem cells but not to those of late passage cells. The phenotype, as well as fat, bone and cartilage differentiation capacity, of mesenchymal stem cells did not change during their culture. Conclusions Extended culture of mesenchymal stem cells alters the ability of these cells to support hematopoietic progenitor cells without causing concomitant changes in their phenotype or differentiation capacity.