Methods for differentiating mesenchymal stem cells
The application discloses a method for obtaining MSC-derived cells with improved transplantation properties from MSC, the method comprising a cell size reduction step, wherein said cell size reduction step is characterized by contacting MSC or MSC-derived cells in vitro or ex vivo with heparin or a derivative or analogue thereof at a concentration of at least 0.01 IU/ml. The application further provides a method for obtaining mesenchymal stem cell-derived cells from mesenchymal stem cells (MSC) comprising contacting MSC in vitro or ex vivo with FGF-2, TGFβ and at least 0.01 IU/ml heparin or a derivative or analogue thereof. The invention also provides the so-obtained cells and cell populations, as well as further products comprising such and uses thereof.
1. A method for obtaining progenitor cells committed to the osteoblastic lineage from mesenchymal stem cells (MSC), the method comprising:
differentiating MSC into progenitor cells committed to osteoblastic lineage by contacting the MSC in vitro or ex vivo with
fibroblast growth factor-2 (FGF-2),
transforming growth factor beta (TGFβ), and
heparin or a derivative or analogue thereof at a concentration of at least 0.01 IU/ml; and
harvesting the progenitor cells committed to the osteoblastic lineage so as to obtain a substantially pure population of the progenitor cells committed to the osteoblastic lineage;
wherein the progenitor cells committed to the osteoblastic lineage have the ability to differentiate into cells of the osteoblastic lineage.
2. The method according to claim 1 , wherein the progenitor cells committed to the osteoblastic lineage are osteoprogenitors or pre-osteoblasts.
3. The method according to claim 1 , wherein the substantially pure population of progenitor cells committed to the osteoblastic lineage comprises at least 90% by number progenitor cells committed to the osteoblastic lineage.
4. The method according to claim 1 , wherein the substantially pure population of progenitor cells committed to the osteoblastic lineage comprises at least 95% by number progenitor cells committed to the osteoblastic lineage.
5. The method according to claim 1 , wherein the the substantially pure population of progenitor cells committed to the osteoblastic lineage comprises progenitor cells committed to the osteoblastic lineage at a concentration between about 1×10 7 /ml and about 1×10 8 /ml of the population.
6. The method according to claim 1 , wherein the progenitor cells committed to the osteoblastic lineage comprise increased expression of a gene encoding an osteoblastic marker selected from the group consisting of Runt-related transcription factor 2 (RUNX2), alkaline phosphatase, biomineralization associated (ALPL), bone morphogenetic protein 2 (BMP2), osteoprotegerin (OPG), periostin (POSTN), cell adhesion molecule 1 (CADM1), connexin 43 (CX43), membrane metalloendopeptidase (CD10), and WNT1 inducible signaling pathway 1 (WISP1) as compared to the expression of the respective gene in MSC.