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Simple Generation of a High Yield Culture of Induced Neurons from Human Adult Skin Fibroblasts

Shrigley, Shelby LU ; Pircs, Karolina LU ; Barker, Roger A. LU ; Parmar, Malin LU and Drouin-ouellet, Janelle LU (2018) In Journal of Visualized Experiments 132.
Abstract
Induced neurons (iNs), the product of somatic cells directly converted to neurons, are a way to obtain patient-derived neurons from tissue thatis easily accessible. Through this route, mature neurons can be obtained in a matter of a few weeks. Here, we describe a straightforward andrapid one-step protocol to obtain iNs from dermal fibroblasts obtained through biopsy samples from adult human donors. We explain each stepof the process, including the maintenance of the dermal fibroblasts, the freezing procedure to build a stock of the cell line, seeding of the cellsfor reprogramming, as well as the culture conditions during the conversion process. In addition, we describe the preparation of glass coverslipsfor electrophysiological recordings,... (More)
Induced neurons (iNs), the product of somatic cells directly converted to neurons, are a way to obtain patient-derived neurons from tissue thatis easily accessible. Through this route, mature neurons can be obtained in a matter of a few weeks. Here, we describe a straightforward andrapid one-step protocol to obtain iNs from dermal fibroblasts obtained through biopsy samples from adult human donors. We explain each stepof the process, including the maintenance of the dermal fibroblasts, the freezing procedure to build a stock of the cell line, seeding of the cellsfor reprogramming, as well as the culture conditions during the conversion process. In addition, we describe the preparation of glass coverslipsfor electrophysiological recordings, long-term coating conditions, and fluorescence activated cell sorting (FACS). We also illustrate examplesof the results to be expected. The protocol described here is easy to perform and can be applied to human fibroblasts derived from human skinbiopsies from patients with various different diagnoses and ages. This protocol generates a sufficient amount of iNs which can be used for a widearray of biomedical applications, including disease modeling, drug screening, and target validation. (Less)
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author
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type
Contribution to journal
publication status
published
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in
Journal of Visualized Experiments
volume
132
publisher
JoVE
ISSN
1940-087X
DOI
10.3791/56904
language
English
LU publication?
yes
id
cd985716-5228-4692-827c-89f68ca47cd6
alternative location
https://www.jove.com/video/56904/simple-generation-high-yield-culture-induced-neurons-from-human-adult
date added to LUP
2018-02-07 09:29:46
date last changed
2018-02-07 14:48:17
@article{cd985716-5228-4692-827c-89f68ca47cd6,
  abstract     = {Induced neurons (iNs), the product of somatic cells directly converted to neurons, are a way to obtain patient-derived neurons from tissue thatis easily accessible. Through this route, mature neurons can be obtained in a matter of a few weeks. Here, we describe a straightforward andrapid one-step protocol to obtain iNs from dermal fibroblasts obtained through biopsy samples from adult human donors. We explain each stepof the process, including the maintenance of the dermal fibroblasts, the freezing procedure to build a stock of the cell line, seeding of the cellsfor reprogramming, as well as the culture conditions during the conversion process. In addition, we describe the preparation of glass coverslipsfor electrophysiological recordings, long-term coating conditions, and fluorescence activated cell sorting (FACS). We also illustrate examplesof the results to be expected. The protocol described here is easy to perform and can be applied to human fibroblasts derived from human skinbiopsies from patients with various different diagnoses and ages. This protocol generates a sufficient amount of iNs which can be used for a widearray of biomedical applications, including disease modeling, drug screening, and target validation.},
  articleno    = {e56904},
  author       = {Shrigley, Shelby and Pircs, Karolina and Barker, Roger A. and Parmar, Malin and Drouin-ouellet, Janelle},
  issn         = {1940-087X},
  language     = {eng},
  month        = {02},
  publisher    = {JoVE},
  series       = {Journal of Visualized Experiments},
  title        = {Simple Generation of a High Yield Culture of Induced Neurons from Human Adult Skin Fibroblasts},
  url          = {http://dx.doi.org/10.3791/56904},
  volume       = {132},
  year         = {2018},
}