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Neuron-Specific Fluorescence Reporter-Based Live Cell Tracing for Transdifferentiation of Mesenchymal Stem Cells into
Do Won Hwang1,2, Hyun Woo Kwon2,3, Jaeho Jang1
1Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul, Republic of Korea.
Stem Cells International
|August 16, 2017
Summary
Mesenchymal stem cells (MSCs) efficiently transform into neurons using the small molecule NHPDQC. A novel fluorescence reporter system allows real-time monitoring of this neuronal differentiation process.
Area of Science:
- Stem Cell Biology
- Neuroscience
- Biochemistry
Background:
- Mesenchymal stem cells (MSCs) hold therapeutic potential for neurodevelopmental disorders.
- Current limitations include inefficient neuronal differentiation and monitoring challenges.
Purpose of the Study:
- To assess the efficacy of NHPDQC in inducing neuronal differentiation of MSCs.
- To develop a method for real-time monitoring of MSC transdifferentiation into neurons.
Main Methods:
- Utilized time-lapse live cell imaging to observe MSCs treated with NHPDQC.
- Employed a red fluorescence reporter plasmid (pTα1-DsRed2) under the tubulin α1 promoter to track neuronal differentiation.
- Quantified neuron-like phenotypes and expression of neuron-specific markers.
Main Results:
- NHPDQC treatment induced neuron-like phenotypes, including neurite outgrowth, in over 95% of MSCs within two days.
- Increased fluorescence signals in reporter-transfected MSCs correlated with neuronal differentiation.
- Real-time monitoring revealed progressive fluorescence increase up to 30 hours, mirroring neurite elongation.
Conclusions:
- NHPDQC is an effective small molecule for efficient MSC neuronal differentiation.
- The developed fluorescence reporter system provides a robust method for monitoring temporal changes in MSC neuronal differentiation.
- This approach facilitates understanding of MSC-to-neuron transdifferentiation and its therapeutic applications.
