Placenta and Bone Marrow-derived MSC Cultured in Serum-free b-FGF-containing Medium Express Cell Surface Frizzled-9 and Give Rise to Multi-Lineage Differentiation

• Published in: Tissue engineering Vol. 13; no. 4; p. 912
• Main Authors: Battula, V L, Bareiss, P, Treml, S, Conrad, S, Albert, I, Hojak, S, Just, L, Skutella, T, Buehring, H-J
• Format: Journal Article
• Language: English
• Published: 01.04.2007
• ISSN: 1076-3279

Background and Aims: Conventionally, MSC are generated by plating bone marrow (BM) cells into culture flasks and selecting plastic-adherent fibroblastoid cells. These cells express high levels of CD9, CD10, CD13, CD73, CD105, CD166, CD318 (CDCP1), and the W8B2 antigen but only low levels of nestin and the embryonic marker SSEA-4. Using a novel protocol we prepared MSC from bone marrow (BM) and non-amniotic placenta (PL) with more immature phenotypes. Methods: Cells from both sources were cultured in gelatine-coated flasks in a serum-free, bFGF-containing medium which is generally used for the maintenance of embryonic stem cells (ESC). Results: Compared to conventionally prepared MSC, BM-MSC generated in ESC medium showed a 4-5 times higher proliferation rate, were positive for frizzled-9 (FZD9) mRNA and surface protein, and expressed higher levels of surface SSEA-4 and Oct-4, nanog-3, and nestin mRNA. In contrast to BM-MSC, PL-MSC were negative for W8B2 antigen but expressed FZD9. Similar to BM-MSC, they expressed increased levels of FZD9 and SSEA-4, when cultured in ESC medium. When MSC from both sources were prepared in ESC medium and cultured under appropriate conditions, they gave rise to adipocytes and osteoblast-like cells (mesoderm), glucagon expressing pancreatic-like cells (endo-derm), as well as neural-like cells expressing the neuronal class III tubulin and the astrocyte marker GFAP (ectoderm). Conclusions: Using a novel culture protocol, we could demonstrate that adult BM- and neonatal placenta-derived MSC can be generated that express a more immature phenotype, show a higher proliferation rate and are capable of multi-potential germ line differentiation.