The morphofunctional properties of induced pluripotent stem cells derived from human skin fibroblasts and differentiated to dopaminergic neurons

Lebedeva, Olga; Lagarkova, M. A.; Sl, Kiselev; Мухина, И.В.; Vedunova, Maria; Usova, O. V.; Stavrovskaya, A. V.; Yamshchikova, N. G.; Fedotova, E. Yu.; Гривенников, И. А.; Хаспеков, Л. Г.; Иллариошкин, С. Н.

doi:10.1134/s1819712413030082

Cited by 8 publications

(8 citation statements)

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“…The subject of our research is mutant Parkin expressed in cells of a patient with a rare PARK2 -associated PD form. The dopaminergic neuron- enriched culture of nerve cells produced by reprogramming of dermal fibroblasts into induced pluripotent stem cells (iPSCs), followed by their directed differentiation, was used as a model system [ 9 , 10 ]. In order to elucidate the disease pathogenesis, we examined the dynamics of several pro- and anti-apoptotic factors in the mutant cell culture in comparison to the neuron culture derived from a healthy donor.…”

Section: Discussionmentioning

confidence: 99%

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Mutations in the Parkinson’s Disease-Associated PARK2 Gene Are Accompanied by Imbalance in Programmed Cell Death Systems

et al. 2015

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Parkinson’s disease is caused by the degeneration of midbrain dopaminergic neurons. A rare recessive form of the disease may be caused by a mutation in the PARK2 gene, whose product, Parkin, controls mitophagy and programmed cell death. The level of pro- and anti-apoptotic factors of the Bcl-2 family was determined in dopaminergic neurons derived from the induced pluripotent stem cells of a healthy donor and a Parkinson’s disease patient bearing PARK2 mutations. Western blotting was used to study the ratios of Bax, Bak, Bcl-2, Bcl-XL, and Bcl-W proteins. The pro-apoptotic Bak protein level in PARK2-neurons was shown to be two times lower than that in healthy cells. In contrast, the expression of the anti-apoptotic factors Bcl-XL, Bcl-W, and Bcl-2 was statistically significantly higher in the mutant cells compared to healthy dopaminergic neurons. These results indicate that PARK2 mutations are accompanied by an imbalance in programmed cell death systems in which non-apoptotic molecular mechanisms play the leading role.

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Section: Discussionmentioning

confidence: 99%

“…The cells were then differentiated into the neuronal type using the differentiation factors. The techniques for producing and re-programming fibroblasts, as well as inducting differentiation of iPSCs into specialized dopaminergic neurons, were described previously [ 10 ].…”

Section: Discussionmentioning

confidence: 99%

Mutations in the Parkinson’s Disease-Associated PARK2 Gene Are Accompanied by Imbalance in Programmed Cell Death Systems

et al. 2015

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“…Lebedeva et al cultured dopamine neurons in vitro and transplanted them into the striatum to improve motor function and alleviate Parkinson's symptoms. [193] They obtained fibroblasts from the skin of three patients with inherited Parkinson's disease with mutations in the LRRK2 and PRKN genes to establish a model of dopaminergic neurons expressing tyrosine hydroxylase on MEA. [193] After 6-OHDA in vitro treatment, the single spikes and bursts of spikes in neural network activity were recorded by MEA in real time, and other neurophysiological status of the cultured piPSC-DN with Parkinson's were monitored.…”

Section: Neural Electrical Activities Of Hipsc-dns Parkinson and Als ...mentioning

confidence: 99%

Using Human‐Induced Pluripotent Stem Cell Derived Neurons on Microelectrode Arrays to Model Neurological Disease: A Review

Lv,

He,

Luo

et al. 2023

Advanced Science

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In situ physiological signals of in vitro neural disease models are essential for studying pathogenesis and drug screening. Currently, an increasing number of in vitro neural disease models are established using human‐induced pluripotent stem cell (hiPSC) derived neurons (hiPSC‐DNs) to overcome interspecific gene expression differences. Microelectrode arrays (MEAs) can be readily interfaced with two‐dimensional (2D), and more recently, three‐dimensional (3D) neural stem cell‐derived in vitro models of the human brain to monitor their physiological activity in real time. Therefore, MEAs are emerging and useful tools to model neurological disorders and disease in vitro using human iPSCs. This is enabling a real‐time window into neuronal signaling at the network scale from patient derived. This paper provides a comprehensive review of MEA's role in analyzing neural disease models established by hiPSC‐DNs. It covers the significance of MEA fabrication, surface structure and modification schemes for hiPSC‐DNs culturing and signal detection. Additionally, this review discusses advances in the development and use of MEA technology to study in vitro neural disease models, including epilepsy, autism spectrum developmental disorder (ASD), and others established using hiPSC‐DNs. The paper also highlights the application of MEAs combined with hiPSC‐DNs in detecting in vitro neurotoxic substances. Finally, the future development and outlook of multifunctional and integrated devices for in vitro medical diagnostics and treatment are discussed.

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“…Human skin fibroblasts were inoculated by 40,000 cells in the medium for fibroblasts with 2 ng/ml bFGF on the 35 mm diameter cultural dish. Two days after expansion the cells were infected with four vectors, obtained previously in our laboratory, for lentiviral expression: LeGO-hOCT4, LeGOhSOX2, LeGO-hc-Myc and LeGO-hKLF4 (MOI -10, 10, 5, 5, respectively) [16,17]. Five days after infecting, the cells were reseeded 1:12 across the area on the culture dishes with 35 mm diameter in the medium for fibroblasts.…”

Section: Methodsmentioning

confidence: 99%

“…New possibilities in modeling PD and other neurodegenerative disorders were discovered in the second half of 2000s as a result of creating the technology of reprogramming of somatic cell to pluripotent stem cells with their further differentiation into the cells of various types, including neurons [14][15][16][17][18][19]. Such reprogramming is realized by the introduction of a certain set of transcription factors into the cell (for example, Oct3/4, Sox2, c-Myc and Klf4).…”

mentioning

confidence: 99%

A Platform for Studying Molecular and Cellular Mechanisms of Parkinson’s Disease Based on Human Induced Pluripotent Stem Cells

Новосадова¹,

Nekrasov²,

Честков³

et al. 2016

Sovrem Tehnol Med

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The aim of the study was to obtain induced pluripotent stem cells (iPSCs) from patients with various forms of Parkinson's disease (PD), and to create on this basis a platform for studying the pathogenesis of the disease at the molecular and cellular level with the development of a protocol of the stem cell differentiation.Materials and Methods. iPSCs were derived from cultured skin fibroblasts, taken from five patients with various forms of PD (PARK8, PARK2, GBA-associated and sporadic forms), and reprogrammed with the help of lentiviral vectors and on the basis of Sendai virus. The obtained iPSCs clones were cultured to the stage of embryonic bodies and, after spontaneous differentiation, stained immunocytochemically. Gene expression and neural markers in these iPSCs lines were analysed using reverse transcription polymerase chain reaction.Results. The obtained iPSCs clones had a normal 46 XY karyotype, stained specifically with Oct4, Nanog, TRA-1-81 and SSEA-4 antibodies, and expressed marker genes responsible for maintaining the pluripotent condition. In the cultures of differentiated iPSCs, cells positively stained for the markers of the three primary germ layers (ectoderm, mesoderm, and endoderm) have been revealed. An effective protocol of iPSCs differentiation into dopaminergic neurons has been worked out, and confirmed by the expression of the specific markertyrosine hydroxylase enzyme.Conclusion. On the basis of explicitly characterized iPSCs from patients with various forms of PD and the developed cellular protocol, a platform for studying the pathogenesis of PD at the molecular and cellular level has been created. Obtaining cell population enriched with dopaminergic neurons opens a perspective for their application for personalized cell replacement PD therapy.

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The morphofunctional properties of induced pluripotent stem cells derived from human skin fibroblasts and differentiated to dopaminergic neurons

Cited by 8 publications

References 23 publications

Mutations in the Parkinson’s Disease-Associated PARK2 Gene Are Accompanied by Imbalance in Programmed Cell Death Systems

Mutations in the Parkinson’s Disease-Associated PARK2 Gene Are Accompanied by Imbalance in Programmed Cell Death Systems

Using Human‐Induced Pluripotent Stem Cell Derived Neurons on Microelectrode Arrays to Model Neurological Disease: A Review

A Platform for Studying Molecular and Cellular Mechanisms of Parkinson’s Disease Based on Human Induced Pluripotent Stem Cells

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