title: Autogenic feeder free system from differentiated mesenchymal progenitor cells, maintains pluripotency of the MEL-1 human embryonic stem cells.
abstract: Human embryonic stem cells (hESc) are known for its pluripotency and self renewal capability, thus possess great potential in regenerative medicine. However, the lack of suitable xenofree extracellular matrix substrate inhibits further applications or the use of hESc in cell-based therapy. In this study, we described a new differentiation method, which generates a homogeneous population of mesenchymal progenitor cells (hESc-MPC) from hESc via epithelial-mesenchymal transition. The extracellular matrix (ECM) proteins from hESc-MPC had in turn supported the undifferentiated expansion of hESc. Immunocytochemistry and flow cytometry characterization of hESc-MPC revealed the presence of early mesenchymal markers. Tandem mass spectometry analysis of ECM produced by hESc-MPC revealed the presence of a mixture of extracellular proteins which includes tenascin C, fibronectin, and vitronectin. The pluripotency of hESc (MEL-1) cultured on the ECM was maintained as shown by the expression of pluripotent genes (FoxD3, Oct-4, Tdgf1, Sox-2, Nanog, hTERT, Rex1), protein markers (SSEA-3, SSEA-4, TRA-1-81, TRA-1-60, Oct-4) and the ability to differentiate into cells representative of ectoderm, endoderm and mesoderm. In summary, we have established a xeno-free autogenic feeder free system to support undifferentiated expansion of hESc, which could be of clinical relevance.
authors.affiliation: UKM Medical Molecular Biology Institute, UKM Medical Center, Jalan Yaacob Latiff, Bandar Tun Razak, 56000 Kuala Lumpur, Malaysia.
authors.forename: Tze Sean
authors.initials: TS
authors.lastname: Khoo
authors.forename: Noor
authors.initials: N
authors.lastname: Hamidah Hussin
authors.forename: Sue-Mian
authors.initials: SM
authors.lastname: Then
authors.forename: Rahman
authors.initials: R
authors.lastname: Jamal
mesh_terms: D002454:Cell Differentiation; D064987:Cell- and Tissue-Based Therapy; D053595:Embryonic Stem Cells / Q000166:cytology*; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics; D005109:Extracellular Matrix / Q000378:metabolism; D061252:Feeder Cells / Q000166:cytology*; D005353:Fibronectins / Q000378:metabolism; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D019063:Tenascin / Q000378:metabolism; D019096:Vitronectin / Q000378:metabolism
chemical_list: D005353:Fibronectins; D019063:Tenascin; D019096:Vitronectin