PMID- 28231034
OWN - NLM
STAT- MEDLINE
DCOM- 20181231
LR  - 20220830
IS  - 2472-5560 (Electronic)
IS  - 2472-5552 (Linking)
VI  - 22
IP  - 3
DP  - 2017 Mar
TI  - Upscaling of hiPS Cell-Derived Neurons for High-Throughput Screening.
PG  - 274-286
LID - 10.1177/1087057116678161 [doi]
AB  - The advent of human-induced pluripotent stem (hiPS) cell-derived neurons promised 
      to provide better model cells for drug discovery in the context of the central 
      nervous system. This work demonstrates both the upscaling of cellular expansion 
      and the acceleration of neuronal differentiation to accommodate the immense 
      material needs of a high-throughput screening (HTS) approach. Using GRowth 
      factor-driven expansion and INhibition of NotCH (GRINCH) during maturation, the 
      derived cells are here referred to as GRINCH neurons. GRINCH cells displayed 
      neuronal markers, and their functional activity could be demonstrated by 
      electrophysiological recordings. In an application of GRINCH neurons, the 
      brain-derived neurotrophic factor (BDNF)-mediated activation of tropomyosin 
      receptor kinase (TrkB) was investigated as a promising drug target to treat 
      synaptic dysfunctions. To assess the phosphorylation of endogenous TrkB in the 
      GRINCH cells, the highly sensitive amplified luminescent proximity homogeneous 
      assay LISA (AlphaLISA) format was established as a primary screen. A 
      high-throughput reverse transcription (RT)-PCR format was employed as a secondary 
      assay to analyze TrkB-mediated downstream target gene expression. In summary, an 
      optimized differentiation protocol, highly efficient cell upscaling, and advanced 
      assay miniaturization, combined with increased detection sensitivity, pave the 
      way for a new generation of predictive cell-based drug discovery.
FAU - Traub, Stefanie
AU  - Traub S
AD  - 1 Lead Identification and Optimization Support, Boehringer Ingelheim Pharma GmbH 
      & Co. KG, Biberach, Germany.
FAU - Stahl, Heiko
AU  - Stahl H
AD  - 2 Immunological and Respiratory Diseases Research, Boehringer Ingelheim Pharma 
      GmbH & Co. KG, Biberach, Germany.
FAU - Rosenbrock, Holger
AU  - Rosenbrock H
AD  - 3 CNS Diseases Research, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, 
      Germany.
FAU - Simon, Eric
AU  - Simon E
AD  - 4 Target Discovery Research, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, 
      Germany.
FAU - Heilker, Ralf
AU  - Heilker R
AD  - 1 Lead Identification and Optimization Support, Boehringer Ingelheim Pharma GmbH 
      & Co. KG, Biberach, Germany.
LA  - eng
PT  - Journal Article
DEP - 20161116
PL  - United States
TA  - SLAS Discov
JT  - SLAS discovery : advancing life sciences R & D
JID - 101697563
RN  - 0 (24-diamino-5-phenylthiazole)
RN  - 0 (Biomarkers)
RN  - 0 (Brain-Derived Neurotrophic Factor)
RN  - 0 (Diamines)
RN  - 0 (Membrane Glycoproteins)
RN  - 0 (Small Molecule Libraries)
RN  - 0 (Thiazoles)
RN  - 103107-01-3 (Fibroblast Growth Factor 2)
RN  - 7171WSG8A2 (BDNF protein, human)
RN  - EC 2.7.10.1 (Receptor, trkB)
RN  - EC 2.7.10.1 (tropomyosin-related kinase-B, human)
SB  - IM
MH  - Animals
MH  - Biomarkers/metabolism
MH  - Brain-Derived Neurotrophic Factor/*genetics/metabolism/pharmacology
MH  - Cell Differentiation
MH  - Cell Proliferation/drug effects
MH  - Diamines/pharmacology
MH  - Drug Discovery/instrumentation/*methods
MH  - Fibroblast Growth Factor 2/pharmacology
MH  - Gene Expression Regulation
MH  - *High-Throughput Screening Assays
MH  - Humans
MH  - Induced Pluripotent Stem Cells/cytology/drug effects/metabolism
MH  - Luminescent Measurements
MH  - Membrane Glycoproteins/*genetics/metabolism
MH  - Neurons/cytology/*drug effects/metabolism
MH  - Patch-Clamp Techniques
MH  - Phosphorylation/drug effects
MH  - Rats
MH  - Receptor, trkB/*genetics/metabolism
MH  - Reverse Transcriptase Polymerase Chain Reaction/instrumentation/methods
MH  - Small Molecule Libraries/*pharmacology
MH  - Thiazoles/pharmacology
OTO - NOTNLM
OT  - human-induced pluripotent stem (hiPS) cells
OT  - neuronal model cells
OT  - tropomyosin receptor kinase B (TrkB)
OT  - upscaling of cell culture
EDAT- 2017/02/24 06:00
MHDA- 2019/01/01 06:00
CRDT- 2017/02/24 06:00
PHST- 2017/02/24 06:00 [entrez]
PHST- 2017/02/24 06:00 [pubmed]
PHST- 2019/01/01 06:00 [medline]
AID - S2472-5552(22)06951-9 [pii]
AID - 10.1177/1087057116678161 [doi]
PST - ppublish
SO  - SLAS Discov. 2017 Mar;22(3):274-286. doi: 10.1177/1087057116678161. Epub 2016 Nov 
      16.