| Title |
A Simplified Method for Generating Purkinje Cells from Human-Induced Pluripotent Stem Cells
|
|---|---|
| Published in |
The Cerebellum, February 2018
|
| DOI | 10.1007/s12311-017-0913-2 |
| Pubmed ID | |
| Authors |
Lauren M. Watson, Maggie M. K. Wong, Jane Vowles, Sally A. Cowley, Esther B. E. Becker |
| Abstract |
The establishment of a reliable model for the study of Purkinje cells in vitro is of particular importance, given their central role in cerebellar function and pathology. Recent advances in induced pluripotent stem cell (iPSC) technology offer the opportunity to generate multiple neuronal subtypes for study in vitro. However, to date, only a handful of studies have generated Purkinje cells from human pluripotent stem cells, with most of these protocols proving challenging to reproduce. Here, we describe a simplified method for the reproducible generation of Purkinje cells from human iPSCs. After 21 days of treatment with factors selected to mimic the self-inductive properties of the isthmic organiser-insulin, fibroblast growth factor 2 (FGF2), and the transforming growth factor β (TGFβ)-receptor blocker SB431542-hiPSCs could be induced to form En1-positive cerebellar progenitors at efficiencies of up to 90%. By day 35 of differentiation, subpopulations of cells representative of the two cerebellar germinal zones, the rhombic lip (Atoh1-positive) and ventricular zone (Ptf1a-positive), could be identified, with the latter giving rise to cells positive for Purkinje cell progenitor-specific markers, including Lhx5, Kirrel2, Olig2 and Skor2. Further maturation was observed following dissociation and co-culture of these cerebellar progenitors with mouse cerebellar cells, with 10% of human cells staining positive for the Purkinje cell marker calbindin by day 70 of differentiation. This protocol, which incorporates modifications designed to enhance cell survival and maturation and improve the ease of handling, should serve to make existing models more accessible, in order to enable future advances in the field. |
X Demographics
The data shown below were collected from the profiles of 13 X users who shared this research output. Click here to find out more about how the information was compiled.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United Kingdom | 2 | 15% |
| Netherlands | 1 | 8% |
| Australia | 1 | 8% |
| Argentina | 1 | 8% |
| Unknown | 8 | 62% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 8 | 62% |
| Scientists | 3 | 23% |
| Practitioners (doctors, other healthcare professionals) | 1 | 8% |
| Science communicators (journalists, bloggers, editors) | 1 | 8% |
Mendeley readers
The data shown below were compiled from readership statistics for 131 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 131 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 29 | 22% |
| Researcher | 24 | 18% |
| Student > Bachelor | 18 | 14% |
| Student > Master | 15 | 11% |
| Student > Postgraduate | 5 | 4% |
| Other | 17 | 13% |
| Unknown | 23 | 18% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 34 | 26% |
| Neuroscience | 30 | 23% |
| Agricultural and Biological Sciences | 20 | 15% |
| Medicine and Dentistry | 7 | 5% |
| Immunology and Microbiology | 3 | 2% |
| Other | 10 | 8% |
| Unknown | 27 | 21% |
Attention Score in Context
This research output has an Altmetric Attention Score of 11. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 24 July 2019.
All research outputs
#3,136,045
of 23,975,976 outputs
Outputs from The Cerebellum
#54
of 957 outputs
Outputs of similar age
#71,674
of 444,986 outputs
Outputs of similar age from The Cerebellum
#3
of 23 outputs
Altmetric has tracked 23,975,976 research outputs across all sources so far. Compared to these this one has done well and is in the 86th percentile: it's in the top 25% of all research outputs ever tracked by Altmetric.
So far Altmetric has tracked 957 research outputs from this source. They receive a mean Attention Score of 3.2. This one has done particularly well, scoring higher than 94% of its peers.
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We're also able to compare this research output to 23 others from the same source and published within six weeks on either side of this one. This one has done well, scoring higher than 86% of its contemporaries.