TY - JOUR

T1 - An Efficient 2D Protocol for Differentiation of iPSCs into Mature Postmitotic Dopaminergic Neurons: Application for Modeling Parkinson’s Disease

AU - Лебедева, О.С.

AU - Шарова, Е.И.

AU - Грехнёв, Дмитрий Александрович

AU - Скородумова, Л.О.

AU - Копылова, И.В.

AU - Вассина, Е.М.

AU - Ошколова, Арина

AU - Новикова, Ю.В.

AU - Крисанова, А.В.

AU - Олехнович, Е.И.

AU - Вигонт, Владимир Александрович

AU - Казначеева, Елена Валентиновна

AU - Богомазова, А.Н.

AU - Лагарькова, М.А.

PY - 2023/4/14

Y1 - 2023/4/14

N2 - About 15% of patients with parkinsonism have a hereditary form of Parkinson's disease (PD). Studies on the early stages of PD pathogenesis are challenging due to the lack of relevant models. The most promising ones are models based on dopaminergic neurons (DAns) differentiated from induced pluripotent stem cells (iPSCs) of patients with hereditary forms of PD. This work describes a highly efficient 2D protocol for obtaining DAns from iPSCs. The protocol is rather simple, comparable in efficiency with previously published protocols, and does not require viral vectors. The resulting neurons have a similar transcriptome profile to previously published data for neurons, and have a high level of maturity marker expression. The proportion of sensitive (SOX6+) DAns in the population calculated from the level of gene expression is higher than resistant (CALB+) DAns. Electrophysiological studies of the DAns confirmed their voltage sensitivity and showed that a mutation in the PARK8 gene is associated with enhanced store-operated calcium entry. The study of high-purity DAns differentiated from the iPSCs of patients with hereditary PD using this differentiation protocol will allow for investigators to combine various research methods, from patch clamp to omics technologies, and maximize information about cell function in normal and pathological conditions.

AB - About 15% of patients with parkinsonism have a hereditary form of Parkinson's disease (PD). Studies on the early stages of PD pathogenesis are challenging due to the lack of relevant models. The most promising ones are models based on dopaminergic neurons (DAns) differentiated from induced pluripotent stem cells (iPSCs) of patients with hereditary forms of PD. This work describes a highly efficient 2D protocol for obtaining DAns from iPSCs. The protocol is rather simple, comparable in efficiency with previously published protocols, and does not require viral vectors. The resulting neurons have a similar transcriptome profile to previously published data for neurons, and have a high level of maturity marker expression. The proportion of sensitive (SOX6+) DAns in the population calculated from the level of gene expression is higher than resistant (CALB+) DAns. Electrophysiological studies of the DAns confirmed their voltage sensitivity and showed that a mutation in the PARK8 gene is associated with enhanced store-operated calcium entry. The study of high-purity DAns differentiated from the iPSCs of patients with hereditary PD using this differentiation protocol will allow for investigators to combine various research methods, from patch clamp to omics technologies, and maximize information about cell function in normal and pathological conditions.

KW - Cell Differentiation/genetics

KW - Dopaminergic Neurons/metabolism

KW - Humans

KW - Induced Pluripotent Stem Cells

KW - Parkinson Disease/metabolism

KW - SOCE currents

KW - iPSCs

KW - calcium

KW - disease modeling

KW - differentiation protocol

KW - high purity of neuron culture

KW - Parkinson’s disease

UR - https://www.mdpi.com/1422-0067/24/8/7297

UR - https://www.mendeley.com/catalogue/11f82679-379a-36d5-99b4-67ca953f56a2/

U2 - 10.3390/ijms24087297

DO - 10.3390/ijms24087297

M3 - Article

C2 - 37108456

VL - 24

JO - International Journal of Molecular Sciences

JF - International Journal of Molecular Sciences

SN - 1422-0067

IS - 8

M1 - 7297

ER -