• Efficient and reproducible generation of MPCs from human ES and iPS cell lines
• Rapid derivation of MPCs in 3 weeks
• Generates MPCs capable of long-term expansion and differentiation to adipocytes, osteoblasts, and chondrocytes
- STEMdiff™-ACF Mesenchymal Induction Medium, 100 mL
- MesenCult™-ACF Basal Medium, 400 mL
- MesenCult™-ACF 5X Supplement, 100 mL
- Animal Component-Free Cell Attachment Substrate, 1 mL
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Data and Publications
Figure 1. Schematic of Differentiation Protocol and Timeline
In Phase 1, human ES or iPS cells are cultured in mTeSR™1 or TeSR™-E8™ medium. On Day 0 (Phase 2) of the protocol, cells are ready for induction into early mesoderm progenitor cells by replacing TeSR™ medium with STEMdiff™ Mesenchymal Induction Medium. By Day 4 (Phase 3), STEMdiff™ Mesenchymal Induction Medium is replaced with MesenCult™-ACF Medium to derive early mesenchymal progenitor cells (MPCs). On Day 6, cells are passaged onto cultureware precoated with MesenCult™-ACF Attachment Substrate in MesenCult™-ACF Medium. By Day 21, human ES- or iPS-derived MPCs exhibit the suggested MPC characteristics.
Figure 2. Cell Expansion and Doubling Rate of MPCs Derived from Human ES (H9) and iPS (STiPS-F016 and -F031) Cells in MesenCult™-ACF Medium
(A) The average cell expansion per passage over 17 passages for MPCs derived from human ES and iPS cell lines are approximately 9 and 10 fold. (B) Days to double cell number for human ES- and iPS-derived MPCs range from 1.1 to 1.4 days.
Figure 3. A Representative Flow Cytometric Analysis of STiPS-F016-derived MPCs Expressing Mesenchymal Surface Markers By Day 21
Human iPS-derived MPCs, generated using the STEMdiff™ Mesenchymal Progenitor Kit, express high levels of mesenchymal surface markers (CD73, CD90 and CD105) and the perivascular marker, CD146. MPCs do not express hematopoietic (CD34, CD45) and endothelial (CD144) surface markers. Human ES-derived MPCs express the same phenotype (data not shown).
Figure 4. Human ES- and iPS-derived MPCs Can Be Further Differentiated Into Adipogenic, Chondrogenic and Osteogenic Lineages
(A) MPCs generated from the 3 week protocol (described in Figure 1) and subsequently cultured in MesenCult™-ACF Medium develop MPC-like morphology (40X magnification). MPCs can be differentiated to (B) adipocytes (Oil Red O staining), 400X magnification; (C) chondrocytes (Alcian Blue staining), 100X magnification; and (D) osteoblasts (Fast Red and Silver Nitrate staining), 100X magnification.