title: Effects of Feeder Cells on Dopaminergic Differentiation of Human Embryonic Stem Cells.
abstract: Mouse embryonic fibroblasts (MEFs) and human foreskin fibroblasts (HFFs) are used for the culture of human embryonic stem cells (hESCs). MEFs and HFFs differed in their capacity to support the proliferation and pluripotency of hESCs and could affect cardiac differentiation potential of hESCs. The aim of this study was to evaluate the effect of MEFs and HFFs feeders on dopaminergic differentiation of hESCs lines. To minimize the impact of culture condition variation, two hESCs lines were cultured on mixed feeder cells (MFCs, MEFs: HFFs = 1:1) and HFFs feeder, respectively, and then were differentiated into dopaminergic (DA) neurons under the identical protocol. Dopaminergic differentiation was evaluated by immunocytochemistry, quantitative fluorescent real-time PCR, transmission and scanning electron microscopy, and patch clamp. Our results demonstrated that these hESCs-derived neurons were genuine and functional DA neurons. However, compared to hESCs line on MFCs feeder, hESCs line on HFFs feeder had a higher proportion of tyrosine hydroxylase (TH) positive cells and expressed higher levels of FOXA2, PITX3, NURR1, and TH genes. In addition, the values of threshold intensity and threshold membrane potential of DA neurons from hESCs line on HFFs feeder were lower than those of DA neurons from hESCs line on the MFCs feeder. In conclusion, HFFs feeder not only facilitated the differentiation of hESCs cells into dopaminergic neurons, but also induced hESCs-derived DA neurons to express higher electrophysiological excitability. Therefore, feeder cells could affect not only dopaminergic differentiation potential of different hESCs lines, but also electrophysiological properties of hESCs-derived DA neurons.
authors.affiliation: Department of Neurology, Jinling Hospital, Southern Medical UniversityNanjing, China; Department of Neurology, First Affiliated Hospital, Hainan Medical UniversityHaikou, China.
authors.forename: Zhenqiang
authors.initials: Z
authors.lastname: Zhao
authors.affiliation: Department of Obstetrics and Gynecology, Nanfang Hospital, Southern Medical UniversityGuangzhou, China; Hainan Provincial Key Laboratory for Human Reproductive Medicine and Genetic Research, Hainan Reproductive Medical Center, First Affiliated Hospital, Hainan Medical UniversityHaikou, China.
authors.forename: Yanlin
authors.initials: Y
authors.lastname: Ma
authors.affiliation: Department of Neurology, First Affiliated Hospital, Hainan Medical University Haikou, China.
authors.forename: Zhibin
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Department of Neurology, Jinling Hospital, Southern Medical University Nanjing, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Liu
authors.affiliation: Hainan Provincial Key Laboratory for Human Reproductive Medicine and Genetic Research, Hainan Reproductive Medical Center, First Affiliated Hospital, Hainan Medical University Haikou, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Li
authors.affiliation: Department of Neurology, Jinling Hospital, Southern Medical UniversityNanjing, China; Department of Neurology, Affiliated Ruikang Hospital, Guangxi Traditional Chinese Medical UniversityNanning, China.
authors.forename: Deyan
authors.initials: D
authors.lastname: Kong
authors.affiliation: Department of Neurology, Jinling Hospital, Southern Medical UniversityNanjing, China; Department of Neurology, Central HospitalShenzhen, China.
authors.forename: Kunxiong
authors.initials: K
authors.lastname: Yuan
authors.affiliation: Department of Laboratory Medicines, First Affiliated Hospital, Hainan Medical University Haikou, China.
authors.forename: Lan
authors.initials: L
authors.lastname: Hu
authors.affiliation: Department of Neurology, First Affiliated Hospital, Hainan Medical University Haikou, China.
authors.forename: Tan
authors.initials: T
authors.lastname: Wang
authors.affiliation: Department of Neurology, First Affiliated Hospital, Hainan Medical University Haikou, China.
authors.forename: Xiaowu
authors.initials: X
authors.lastname: Chen
authors.affiliation: Department of Neurology, First Affiliated Hospital, Hainan Medical University Haikou, China.
authors.forename: Yanan
authors.initials: Y
authors.lastname: Peng
authors.affiliation: Hainan Provincial Key Laboratory for Human Reproductive Medicine and Genetic Research, Hainan Reproductive Medical Center, First Affiliated Hospital, Hainan Medical University Haikou, China.
authors.forename: Weimin
authors.initials: W
authors.lastname: Jiang
authors.affiliation: Department of Obstetrics and Gynecology, Nanfang Hospital, Southern Medical University Guangzhou, China.
authors.forename: Yanhong
authors.initials: Y
authors.lastname: Yu
authors.affiliation: Department of Neurology, Jinling Hospital, Southern Medical University Nanjing, China.
authors.forename: Xinfeng
authors.initials: X
authors.lastname: Liu
keywords: dopaminergic neuron; electrophysiological properties; human embryonic stem cells; human foreskin fibroblasts; mouse embryonic fibroblasts; neural differentiation