Background: Human ES cells can proliferate without a known limit and can form advanced derivatives of all three embryonic germ layers. The potential of human embryonic stem (hES) cells to differentiate into cell types of a variety of organs has generated much excitement over the possible use of hES cells in therapeutic applications. Thus, it becomes crucial to develop protocols for the directed differentiation of embryonic stem cells into tissue-restricted precursors. Here, we present optimal culture conditions for the derivation of unlimited numbers of pure mesenchymal precursors from human embryonic stem cells.
Methods and results: We evaluated efficiency of various cytokine based commitment condition of hES cell into multipotent mesenchymal precursors (MMPs). And we evaluated changes of surface maker expression of hES after cytokine treatment and multi-lineage differentiation capacity of hES derived MMPs. We can induce differentiation of hES cell into homogenous spindle shaped cells after 5 weeks incubation with BMP based conditioned media. hES derived spindle shaped cells (MMPs) highly expressed SH2(CD105), SH3(CD73) and ICAM -1(CD54) but rarely expressed hES cell markers such as SSEA-4 and TRA-1-81. Purity of hES derived MMPs is significantly higher than previous report. hES derived MMPs displayed a stable phenotype up to 40 passages and remained as a monolayer in vitro. hES derived MMPs could be induced to differentiate exclusively into the adipocytic, chondrocytic, osteocytic and myogenic lineages.
Conclusion: Our findings will help to elucidate the mechanism of mesoderm specification during embryonic stem cell differentiation and provide a platform to efficiently generate specialized human mesenchymal cell types for future clinical applications.
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