Selection via pluripotency-related transcriptional screen minimizes the influence of somatic origin on iPSC differentiation propensity

Katherine A. Hartjes, Xing Li, Almudena Martinez-Fernandez, Alexa J. Roemmich, Brandon Larsen, Andre Terzic, Timothy J Nelson

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

The value of induced pluripotent stem cells (iPSCs) within regenerative medicine is contingent on predictable and consistent iPSC differentiation. However, residual influence of the somatic origin or reprogramming technique may variegate differentiation propensity and confound comparative genotype/phenotype analyses. The objective of this study was to define quality control measures to select iPSC clones that minimize the influence of somatic origin on differentiation propensity independent of the reprogramming strategy. More than 60 murine iPSC lines were derived from different fibroblast origins (embryonic, cardiac, and tail tip) via lentiviral integration and doxycycline-induced transgene expression. Despite apparent equivalency according to established iPSC histologic and cytomorphologic criteria, clustering of clonal variability in pluripotency-related gene expression identified transcriptional outliers that highlighted cell lines with unpredictable cardiogenic propensity. Following selection according to a standardized gene expression profile calibrated by embryonic stem cells, the influence of somatic origin on iPSC methylation and transcriptional patterns was negated. Furthermore, doxycycline-induced iPSCs consistently demonstrated earlier differentiation than lentiviral-reprogrammed lines using contractile cardiac tissue as a measure of functional differentiation. Moreover, delayed cardiac differentiation was predominately associated with upregulation in pluripotency-related gene expression upon differentiation. Starting from a standardized pool of iPSCs, relative expression levels of two pluripotency genes, Oct4 and Zfp42, statistically correlated with enhanced cardiogenicity independent of somatic origin or reprogramming strategy (R2 = 0.85). These studies demonstrate that predictable iPSC differentiation is independent of somatic origin with standardized gene expression selection criteria, while the residual impact of reprogramming strategy greatly influences predictable output of tissue-specification required for comparative genotype/phenotype analyses. Stem Cells 2014;32:2350-2359

Original languageEnglish (US)
Pages (from-to)2350-2359
Number of pages10
JournalStem Cells
Volume32
Issue number9
DOIs
StatePublished - 2014

Fingerprint

Induced Pluripotent Stem Cells
Cell Differentiation
Doxycycline
Gene Expression
Genotype
Phenotype
Cell Line
Regenerative Medicine
Embryonic Stem Cells
Transgenes
Transcriptome
Quality Control
Patient Selection
Methylation
Cluster Analysis
Up-Regulation
Stem Cells
Clone Cells
Fibroblasts

Keywords

  • Cardiac differentiation
  • Induced pluripotent stem cells
  • Transcriptional profile

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Molecular Medicine

Cite this

Selection via pluripotency-related transcriptional screen minimizes the influence of somatic origin on iPSC differentiation propensity. / Hartjes, Katherine A.; Li, Xing; Martinez-Fernandez, Almudena; Roemmich, Alexa J.; Larsen, Brandon; Terzic, Andre; Nelson, Timothy J.

In: Stem Cells, Vol. 32, No. 9, 2014, p. 2350-2359.

Research output: Contribution to journalArticle

@article{a5491176b1ea4658862ebeb5413610bf,
title = "Selection via pluripotency-related transcriptional screen minimizes the influence of somatic origin on iPSC differentiation propensity",
abstract = "The value of induced pluripotent stem cells (iPSCs) within regenerative medicine is contingent on predictable and consistent iPSC differentiation. However, residual influence of the somatic origin or reprogramming technique may variegate differentiation propensity and confound comparative genotype/phenotype analyses. The objective of this study was to define quality control measures to select iPSC clones that minimize the influence of somatic origin on differentiation propensity independent of the reprogramming strategy. More than 60 murine iPSC lines were derived from different fibroblast origins (embryonic, cardiac, and tail tip) via lentiviral integration and doxycycline-induced transgene expression. Despite apparent equivalency according to established iPSC histologic and cytomorphologic criteria, clustering of clonal variability in pluripotency-related gene expression identified transcriptional outliers that highlighted cell lines with unpredictable cardiogenic propensity. Following selection according to a standardized gene expression profile calibrated by embryonic stem cells, the influence of somatic origin on iPSC methylation and transcriptional patterns was negated. Furthermore, doxycycline-induced iPSCs consistently demonstrated earlier differentiation than lentiviral-reprogrammed lines using contractile cardiac tissue as a measure of functional differentiation. Moreover, delayed cardiac differentiation was predominately associated with upregulation in pluripotency-related gene expression upon differentiation. Starting from a standardized pool of iPSCs, relative expression levels of two pluripotency genes, Oct4 and Zfp42, statistically correlated with enhanced cardiogenicity independent of somatic origin or reprogramming strategy (R2 = 0.85). These studies demonstrate that predictable iPSC differentiation is independent of somatic origin with standardized gene expression selection criteria, while the residual impact of reprogramming strategy greatly influences predictable output of tissue-specification required for comparative genotype/phenotype analyses. Stem Cells 2014;32:2350-2359",
keywords = "Cardiac differentiation, Induced pluripotent stem cells, Transcriptional profile",
author = "Hartjes, {Katherine A.} and Xing Li and Almudena Martinez-Fernandez and Roemmich, {Alexa J.} and Brandon Larsen and Andre Terzic and Nelson, {Timothy J}",
year = "2014",
doi = "10.1002/stem.1734",
language = "English (US)",
volume = "32",
pages = "2350--2359",
journal = "Stem Cells",
issn = "1066-5099",
publisher = "Wiley-Blackwell",
number = "9",

}

TY - JOUR

T1 - Selection via pluripotency-related transcriptional screen minimizes the influence of somatic origin on iPSC differentiation propensity

AU - Hartjes, Katherine A.

AU - Li, Xing

AU - Martinez-Fernandez, Almudena

AU - Roemmich, Alexa J.

AU - Larsen, Brandon

AU - Terzic, Andre

AU - Nelson, Timothy J

PY - 2014

Y1 - 2014

N2 - The value of induced pluripotent stem cells (iPSCs) within regenerative medicine is contingent on predictable and consistent iPSC differentiation. However, residual influence of the somatic origin or reprogramming technique may variegate differentiation propensity and confound comparative genotype/phenotype analyses. The objective of this study was to define quality control measures to select iPSC clones that minimize the influence of somatic origin on differentiation propensity independent of the reprogramming strategy. More than 60 murine iPSC lines were derived from different fibroblast origins (embryonic, cardiac, and tail tip) via lentiviral integration and doxycycline-induced transgene expression. Despite apparent equivalency according to established iPSC histologic and cytomorphologic criteria, clustering of clonal variability in pluripotency-related gene expression identified transcriptional outliers that highlighted cell lines with unpredictable cardiogenic propensity. Following selection according to a standardized gene expression profile calibrated by embryonic stem cells, the influence of somatic origin on iPSC methylation and transcriptional patterns was negated. Furthermore, doxycycline-induced iPSCs consistently demonstrated earlier differentiation than lentiviral-reprogrammed lines using contractile cardiac tissue as a measure of functional differentiation. Moreover, delayed cardiac differentiation was predominately associated with upregulation in pluripotency-related gene expression upon differentiation. Starting from a standardized pool of iPSCs, relative expression levels of two pluripotency genes, Oct4 and Zfp42, statistically correlated with enhanced cardiogenicity independent of somatic origin or reprogramming strategy (R2 = 0.85). These studies demonstrate that predictable iPSC differentiation is independent of somatic origin with standardized gene expression selection criteria, while the residual impact of reprogramming strategy greatly influences predictable output of tissue-specification required for comparative genotype/phenotype analyses. Stem Cells 2014;32:2350-2359

AB - The value of induced pluripotent stem cells (iPSCs) within regenerative medicine is contingent on predictable and consistent iPSC differentiation. However, residual influence of the somatic origin or reprogramming technique may variegate differentiation propensity and confound comparative genotype/phenotype analyses. The objective of this study was to define quality control measures to select iPSC clones that minimize the influence of somatic origin on differentiation propensity independent of the reprogramming strategy. More than 60 murine iPSC lines were derived from different fibroblast origins (embryonic, cardiac, and tail tip) via lentiviral integration and doxycycline-induced transgene expression. Despite apparent equivalency according to established iPSC histologic and cytomorphologic criteria, clustering of clonal variability in pluripotency-related gene expression identified transcriptional outliers that highlighted cell lines with unpredictable cardiogenic propensity. Following selection according to a standardized gene expression profile calibrated by embryonic stem cells, the influence of somatic origin on iPSC methylation and transcriptional patterns was negated. Furthermore, doxycycline-induced iPSCs consistently demonstrated earlier differentiation than lentiviral-reprogrammed lines using contractile cardiac tissue as a measure of functional differentiation. Moreover, delayed cardiac differentiation was predominately associated with upregulation in pluripotency-related gene expression upon differentiation. Starting from a standardized pool of iPSCs, relative expression levels of two pluripotency genes, Oct4 and Zfp42, statistically correlated with enhanced cardiogenicity independent of somatic origin or reprogramming strategy (R2 = 0.85). These studies demonstrate that predictable iPSC differentiation is independent of somatic origin with standardized gene expression selection criteria, while the residual impact of reprogramming strategy greatly influences predictable output of tissue-specification required for comparative genotype/phenotype analyses. Stem Cells 2014;32:2350-2359

KW - Cardiac differentiation

KW - Induced pluripotent stem cells

KW - Transcriptional profile

UR - http://www.scopus.com/inward/record.url?scp=84906242477&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84906242477&partnerID=8YFLogxK

U2 - 10.1002/stem.1734

DO - 10.1002/stem.1734

M3 - Article

VL - 32

SP - 2350

EP - 2359

JO - Stem Cells

JF - Stem Cells

SN - 1066-5099

IS - 9

ER -