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1.
PLoS One ; 3(8): e2930, 2008 Aug 13.
Article in English | MEDLINE | ID: mdl-18698373

ABSTRACT

Wnt signals exercise strong cell-biological and regenerative effects of considerable therapeutic value. There are, however, no specific Wnt agonists and no method for in vivo delivery of purified Wnt proteins. Wnts contain lipid adducts that are required for activity and we exploited this lipophilicity by packaging purified Wnt3a protein into lipid vesicles. Rather than being encapsulated, Wnts are tethered to the liposomal surface, where they enhance and sustain Wnt signaling in vitro. Molecules that effectively antagonize soluble Wnt3a protein but are ineffective against the Wnt3a signal presented by a cell in a paracrine or autocrine manner are also unable to block liposomal Wnt3a activity, suggesting that liposomal packaging mimics the biological state of active Wnts. When delivered subcutaneously, Wnt3a liposomes induce hair follicle neogenesis, demonstrating their robust biological activity in a regenerative context.


Subject(s)
Wnt Proteins/biosynthesis , Wnt Proteins/physiology , Animals , Cell Culture Techniques , Liposomes , Male , Membrane Lipids/physiology , Mice , Mice, Inbred Strains , Signal Transduction , Wnt Proteins/antagonists & inhibitors , Wnt Proteins/isolation & purification , Wnt3 Protein , Wnt3A Protein
2.
Hum Reprod ; 19(12): 2875-83, 2004 Dec.
Article in English | MEDLINE | ID: mdl-15375076

ABSTRACT

BACKGROUND: The aim of this study was to characterize human embryonic stem (ES) cells at the molecular level by performing large-scale complementary DNA (cDNA) analysis using DNA micro-arrays. METHODS: The transcription profile of human ES cells was determined by comparing it to 2, 10 and 30-day old embryoid bodies (EBs) using Affymetrix Genechip human micro-arrays (U133). RESULTS: According to this analysis we demonstrate that two human ES cell lines are more close to each other than to their differentiated derivatives. We also show the spectrum of cytokine receptors that they express, and demonstrate the presence of five genes that are highly specific to human ES cells and to germ cells. Moreover, by profiling different stages in the differentiation of human embryoid bodies, we illustrate the clustering of five sets of temporally expressed genes, which could be related to the sequential stages of embryonic development. Among them are known genes that are involved in early pattern formation. CONCLUSIONS: The present study provides a molecular basis for the identity of human ES cells and demonstrates that during their in vitro differentiation they express embryonic specific genes in a stage specific manner.


Subject(s)
Cell Differentiation/genetics , Embryo, Mammalian/cytology , Gene Expression , Stem Cells/physiology , Cells, Cultured , DNA-Binding Proteins/genetics , Homeodomain Proteins/genetics , Humans , Left-Right Determination Factors , Nodal Protein , Octamer Transcription Factor-3 , Oligonucleotide Array Sequence Analysis , Pulmonary Surfactant-Associated Protein D/genetics , Transcription Factors/genetics , Transforming Growth Factor beta/genetics , alpha-Fetoproteins/genetics , Homeobox Protein PITX2
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