Evidence map›Paper›PMID 29024560›Full record

ArticleStem cells translational medicine2017

Enhanced Stem Cell Differentiation and Immunopurification of Genome Engineered Human Retinal Ganglion Cells.

Valentin M Sluch, Xitiz Chamling, Melissa M Liu, Cynthia A Berlinicke, Jie Cheng, Katherine L Mitchell, Derek S Welsbie, Donald J Zack

Open access · goldAbstract read
In one paragraph

Article in Stem cells translational medicine, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 100 papers.

0numbers the graph read from it
0cells of the map it votes in
100citing papers in PubMed
4.5field-weighted citation impact, top 4% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

100 citing papers in PubMed, 140 citations in OpenAlex.

  1. Article
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  4. Neuroinflammation in glaucoma: a myriad of cellular pathways and players.Mammalian genome : official journal of the International Mammalian Genome Society · 2026
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  14. Single-Cell Transcriptomics onBiomedicines · 2025
    Article
  15. Review
  16. Article
  17. Dynamic changes in mRNA isoform usage during human retinal development.bioRxiv : the preprint server for biology · 2025
    Article
  18. Article
  19. Article
  20. CUT&TIME captures the history of open chromatin in developing neurons.bioRxiv : the preprint server for biology · 2025
    Article

40 more citing papers are in PubMed but not listed here.

4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

8 authors at 3 institutions in 1 country.

Valentin M SluchDepartment of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.ORCID 0000-0003-3318-7986
Xitiz ChamlingDepartment of Ophthalmology, Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
Melissa M LiuDepartment of Ophthalmology, Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
Cynthia A BerlinickeDepartment of Ophthalmology, Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
Jie ChengDepartment of Ophthalmology, Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
Katherine L MitchellDepartment of Ophthalmology, Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
Derek S WelsbieDepartment of Ophthalmology, Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
Donald J ZackDepartment of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
Johns Hopkins Medicine · USJohns Hopkins University · USSmith-Kettlewell Eye Research Institute · US

Funding

MEDICAL SCIENTIST TRAINING PROGRAMT32GM007309 · NIGMS · JOHNS HOPKINS UNIVERSITY · PI COX, ANDREA L · 1985 to 2019
$43.2M
Wilmer Core Grant for Vision ResearchP30EY001765 · NEI · JOHNS HOPKINS UNIVERSITY · PI Jiang Qian · 1985 to 2026
$22.1M
Visual Science Training ProgramT32EY007143 · NEI · JOHNS HOPKINS UNIVERSITY · PI Laura Ensign, JEREMY NATHANS · 1995 to 2026
$6.2M
A High Content Screening Approach for the Retinal Degenerative DiseasesR01EY024249 · NEI · JOHNS HOPKINS UNIVERSITY · PI BERLINICKE, CYNTHIA · 2015 to 2019
$2.0M
Dual Leucine Zipper Kinase (DLK) as a Mediator of Retinal Ganglion Cell InjuryR01EY023754 · NEI · JOHNS HOPKINS UNIVERSITY · PI ZACK, DONALD J. · 2013 to 2016
$1.6M
NEI NIH HHS P30 EY001765NEI NIH HHS R01 EY023754NEI NIH HHS R01 EY024249NEI NIH HHS T32 EY007143NIGMS NIH HHS T32 GM007309
6 · The paper itself

Abstract

Human pluripotent stem cells have the potential to promote biological studies and accelerate drug discovery efforts by making possible direct experimentation on a variety of human cell types of interest. However, stem cell cultures are generally heterogeneous and efficient differentiation and purification protocols are often lacking. Here, we describe the generation of clustered regularly-interspaced short palindromic repeats(CRISPR)-Cas9 engineered reporter knock-in embryonic stem cell lines in which tdTomato and a unique cell-surface protein, THY1.2, are expressed under the control of the retinal ganglion cell (RGC)-enriched gene BRN3B. Using these reporter cell lines, we greatly improved adherent stem cell differentiation to the RGC lineage by optimizing a novel combination of small molecules and established an anti-THY1.2-based protocol that allows for large-scale RGC immunopurification. RNA-sequencing confirmed the similarity of the stem cell-derived RGCs to their endogenous human counterparts. Additionally, we developed an in vitro axonal injury model suitable for studying signaling pathways and mechanisms of human RGC cell death and for high-throughput screening for neuroprotective compounds. Using this system in combination with RNAi-based knockdown, we show that knockdown of dual leucine kinase (DLK) promotes survival of human RGCs, expanding to the human system prior reports that DLK inhibition is neuroprotective for murine RGCs. These improvements will facilitate the development and use of large-scale experimental paradigms that require numbers of pure RGCs that were not previously obtainable. Stem Cells Translational Medicine 2017;6:1972-1986.

Indexed as

Cell DifferentiationCell LineCells, CulturedCellular Reprogramming TechniquesCRISPR-Cas SystemsGene EditingHuman Embryonic Stem CellsHumansMAP Kinase Kinase KinasesRetinal Ganglion CellsTranscription Factor Brn-3BMAP Kinase Kinase Kinasesmitogen-activated protein kinase kinase kinase 12POU4F2 protein, humanTranscription Factor Brn-3BBiotechnologyCell differentiationClustered regularly interspaced short palindromic repeatsRetinal ganglion cellsStem cells

Identifiers

PMID29024560
PMCPMC6430043
OpenAlexW2761739377

What OpenQuestion holds

Textmetadata
LicenceCC BY-NC-ND
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.