Anita Laitinen1, Milla Lampinen2, Stefanie Liedtke3, Lotta Kilpinen4, Erja Kerkelä4, Jertta-Riina Sarkanen5, Tuula Heinonen6, Gesine Kogler3, Saara Laitinen7. 1. Cell Therapy Services, Medical Services, Finnish Red Cross Blood Service, Helsinki, Finland; Research and Development, Medical Services, Finnish Red Cross Blood Service, Helsinki, Finland. 2. Faculty of Medicine, Department of Pharmacology, University of Helsinki, Helsinki, Finland; Research and Development, Medical Services, Finnish Red Cross Blood Service, Helsinki, Finland. 3. Institute of Transplantation Diagnostics and Cell Therapeutics, Heinrich-Heine-University Medical Center, Düsseldorf, Germany. 4. Research and Development, Medical Services, Finnish Red Cross Blood Service, Helsinki, Finland. 5. FICAM Finnish Center for Alternative Methods, School of Medicine, University of Tampere, Tampere, Finland; Cell Biology, School of Medicine, University of Tampere, Tampere, Finland. 6. FICAM Finnish Center for Alternative Methods, School of Medicine, University of Tampere, Tampere, Finland. 7. Research and Development, Medical Services, Finnish Red Cross Blood Service, Helsinki, Finland. Electronic address: saara.laitinen@bloodservice.fi.
Abstract
BACKGROUND AIMS: Cord blood (CB) is an attractive source of mesenchymal stromal cells (MSCs) because of its abundant availability and ease of collection. However, the success rate of generating CB-MSCs is low. In this study, our aim was to demonstrate the efficiency of our previously described method to obtain MSCs from CB and further characterize them and to study the effects of different culture conditions on MSCs. METHODS: CB-MSC cultures were established in low oxygen (3%) conditions on fibronectin in 10% fetal bovine serum containing culture medium supplemented with combinations of growth factors. Cells were characterized for their adipogenic, osteogenic and chondrogenic differentiation capacity; phenotype; and HOX gene expression profile. The functionality of the cells cultured in different media was tested in vitro with angiogenesis and T-cell proliferation assays. RESULTS: We demonstrate 87% efficacy in generating MSCs from CB. The established cells had typical MSC characteristics with reduced adipogenic differentiation potential and a unique HOX gene fingerprint. Growth factor-rich medium and a 3% oxygen condition enhanced cell proliferation; however, the growth factor-rich medium had a negative effect on the expression of CD90. Dexamethasone-containing medium improved the capacity of the cells to suppress T-cell proliferation, whereas the cells grown without dexamethasone were more able to support angiogenesis. CONCLUSIONS: Our results demonstrate that the composition of expansion medium is critical for the functionality of MSCs and should always be appropriately defined for each purpose.
BACKGROUND AIMS: Cord blood (CB) is an attractive source of mesenchymal stromal cells (MSCs) because of its abundant availability and ease of collection. However, the success rate of generating CB-MSCs is low. In this study, our aim was to demonstrate the efficiency of our previously described method to obtain MSCs from CB and further characterize them and to study the effects of different culture conditions on MSCs. METHODS: CB-MSC cultures were established in low oxygen (3%) conditions on fibronectin in 10% fetal bovine serum containing culture medium supplemented with combinations of growth factors. Cells were characterized for their adipogenic, osteogenic and chondrogenic differentiation capacity; phenotype; and HOX gene expression profile. The functionality of the cells cultured in different media was tested in vitro with angiogenesis and T-cell proliferation assays. RESULTS: We demonstrate 87% efficacy in generating MSCs from CB. The established cells had typical MSC characteristics with reduced adipogenic differentiation potential and a unique HOX gene fingerprint. Growth factor-rich medium and a 3% oxygen condition enhanced cell proliferation; however, the growth factor-rich medium had a negative effect on the expression of CD90. Dexamethasone-containing medium improved the capacity of the cells to suppress T-cell proliferation, whereas the cells grown without dexamethasone were more able to support angiogenesis. CONCLUSIONS: Our results demonstrate that the composition of expansion medium is critical for the functionality of MSCs and should always be appropriately defined for each purpose.