Evidence map›Paper›PMID 25286293›Full record

ArticleJournal of visualized experiments : JoVE2014

High efficiency differentiation of human pluripotent stem cells to cardiomyocytes and characterization by flow cytometry.

Subarna Bhattacharya, Paul W Burridge, Erin M Kropp, Sandra L Chuppa, Wai-Meng Kwok, Joseph C Wu, Kenneth R Boheler, Rebekah L Gundry

Open access · bronzeAbstract readVideo-Audio Media
In one paragraph

Article in Journal of visualized experiments : JoVE, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 55 papers, 1 of them a synthesis that pooled it.

0numbers the graph read from it
0cells of the map it votes in
55citing papers in PubMed, 1 pooled it
3.1field-weighted citation impact, top 8% 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

55 citing papers in PubMed, 1 synthesis or guideline pooled it, 82 citations in OpenAlex.

  1. Pooled it
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  6. Review
  7. Environmental Cues Facilitate Maturation and Patterning of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes.Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology · 2024
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  16. Personalized Medicine Approach in a DCM Patient withJournal of personalized medicine · 2022
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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 4 institutions in 2 countries.

Subarna BhattacharyaDepartment of Biochemistry, Medical College of Wisconsin.
Paul W BurridgeStanford Cardiovascular Institute, Stanford University School of Medicine.
Erin M KroppDepartment of Biochemistry, Medical College of Wisconsin.
Sandra L ChuppaDepartment of Biochemistry, Medical College of Wisconsin.
Wai-Meng KwokDepartment of Anesthesiology, Medical College of Wisconsin.
Joseph C WuStanford Cardiovascular Institute, Stanford University School of Medicine.
Kenneth R BohelerStem Cell and Regenerative Medicine Consortium, LKS Faculty of Medicine, Hong Kong University; Division of Cardiology, Johns Hopkins University School of Medicine.
Rebekah L GundryDepartment of Biochemistry, Medical College of Wisconsin; Cardiovascular Research Center, Biotechnology and Bioengineering Center, Medical College of Wisconsin; rgundry@mcw.edu.
Medical College of Wisconsin · USCardiovascular Institute of the South · USJohns Hopkins Medicine · USStanford University · US

Funding

Induced pluripotent stem cells in the understanding and treatment of heart diseaseU01HL099776 · NHLBI · STANFORD UNIVERSITY · PI LONGAKER, MICHAEL T, WU, JOSEPH C. · 2009 to 2015
$8.2M
Surface Proteins in the Differentiation of Stem Cells to CardiomyocytesR00HL094708 · NHLBI · MEDICAL COLLEGE OF WISCONSIN · PI GUNDRY, REBEKAH L. · 2011 to 2013
$702k
NHLBI NIH HHS 4R00HL094708-03NHLBI NIH HHS R00 HL094708NHLBI NIH HHS U01 HL099776
6 · The paper itself

Abstract

There is an urgent need to develop approaches for repairing the damaged heart, discovering new therapeutic drugs that do not have toxic effects on the heart, and improving strategies to accurately model heart disease. The potential of exploiting human induced pluripotent stem cell (hiPSC) technology to generate cardiac muscle "in a dish" for these applications continues to generate high enthusiasm. In recent years, the ability to efficiently generate cardiomyogenic cells from human pluripotent stem cells (hPSCs) has greatly improved, offering us new opportunities to model very early stages of human cardiac development not otherwise accessible. In contrast to many previous methods, the cardiomyocyte differentiation protocol described here does not require cell aggregation or the addition of Activin A or BMP4 and robustly generates cultures of cells that are highly positive for cardiac troponin I and T (TNNI3, TNNT2), iroquois-class homeodomain protein IRX-4 (IRX4), myosin regulatory light chain 2, ventricular/cardiac muscle isoform (MLC2v) and myosin regulatory light chain 2, atrial isoform (MLC2a) by day 10 across all human embryonic stem cell (hESC) and hiPSC lines tested to date. Cells can be passaged and maintained for more than 90 days in culture. The strategy is technically simple to implement and cost-effective. Characterization of cardiomyocytes derived from pluripotent cells often includes the analysis of reference markers, both at the mRNA and protein level. For protein analysis, flow cytometry is a powerful analytical tool for assessing quality of cells in culture and determining subpopulation homogeneity. However, technical variation in sample preparation can significantly affect quality of flow cytometry data. Thus, standardization of staining protocols should facilitate comparisons among various differentiation strategies. Accordingly, optimized staining protocols for the analysis of IRX4, MLC2v, MLC2a, TNNI3, and TNNT2 by flow cytometry are described.

Indexed as

Cell DifferentiationEmbryonic Stem CellsFlow CytometryHomeodomain ProteinsHumansInduced Pluripotent Stem CellsMyocytes, CardiacPluripotent Stem CellsHomeodomain ProteinsIRX4 protein, human

Identifiers

PMID25286293
PMCPMC4448667
OpenAlexW2014131661

What OpenQuestion holds

Textmetadata
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.