title: Alginate microcapsule as a 3D platform for the efficient differentiation of human embryonic stem cells to dopamine neurons.
abstract: Human embryonic stem cells (hESCs) are emerging as an attractive alternative source for cell replacement therapy since the cells can be expanded in culture indefinitely and differentiated into any cell types in the body. In order to optimize cell-to-cell interaction, cell proliferation and differentiation into specific lineages as well as tissue organization, it is important to provide a microenvironment for the hESCs which mimics the stem cell niche. One approach is to provide a three-dimensional (3D) environment such as encapsulation. We present an approach to culture and differentiate hESCs into midbrain dopamine (mdDA) neurons in a 3D microenvironment using alginate microcapsules for the first time. A detailed gene and protein expression analysis during neuronal differentiation showed an increased gene and protein expression of various specific DA neuronal markers, particularly tyrosine hydroxylase (TH) by >100 folds after 2 weeks and at least 50% higher expression after 4 weeks respectively, compared to cells differentiated under conventional two-dimensional (2D) platform. The encapsulated TH(+) cells co-expressed mdDA neuronal markers, forkhead box protein A-2 (FOXA2) and pituitary homeobox-3 (PITX3) after 4 weeks and secreted approximately 60pg/ml/10(6) cells higher DA level when induced. We propose that the 3D platform facilitated an early onset of DA neuronal generation compared to that with conventional 2D system which also secretes more DA under potassium-induction. It is a very useful model to study the proliferation and directed differentiation of hESCs to various lineages, particularly to mdDA neurons. This 3D system also allows the separation of feeder cells from hESCs during the process of differentiation and also has potential for immune-isolation during transplantation studies.
authors.affiliation: Stem Cell Lab, Faculty of Medicine, School of Psychiatry, University of New South Wales, Australia. Electronic address: jaeminkim85@hotmail.com.
authors.forename: Jaemin
authors.initials: J
authors.lastname: Kim
authors.forename: Perminder
authors.initials: P
authors.lastname: Sachdev
authors.forename: Kuldip
authors.initials: K
authors.lastname: Sidhu
mesh_terms: D000464:Alginates / Q000737:chemistry*; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002214:Capsules / Q000737:chemistry*; D018929:Cell Culture Techniques; D002460:Cell Line; D019070:Cell Lineage; D049109:Cell Proliferation; D018920:Coculture Techniques; D004298:Dopamine / Q000378:metabolism; D059290:Dopaminergic Neurons / Q000166:cytology* / Q000201:enzymology / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology*; D020723:Glucuronic Acid / Q000737:chemistry; D051542:Hepatocyte Nuclear Factor 3-beta / Q000378:metabolism; D006603:Hexuronic Acids / Q000737:chemistry; D018398:Homeodomain Proteins / Q000378:metabolism; D006801:Humans; D051379:Mice; D008954:Models, Biological; D055495:Neurogenesis; D014157:Transcription Factors / Q000378:metabolism; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism
chemical_list: D000464:Alginates; D015415:Biomarkers; D002214:Capsules; C496502:FOXA2 protein, human; D006603:Hexuronic Acids; D018398:Homeodomain Proteins; D014157:Transcription Factors; C433629:homeobox protein PITX3; D051542:Hepatocyte Nuclear Factor 3-beta; D020723:Glucuronic Acid; D014446:Tyrosine 3-Monooxygenase; D004298:Dopamine