Marmoset induced pluripotent stem cells: Robust neural differentiation following pretreatment with dimethyl sulfoxide

Zhifang Qiu, Anuja Mishra, Miao Li, Steven L. Farnsworth, Bernadette Guerra, Robert E. Lanford, Peter J Hornsby

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

The marmoset is an important nonhuman primate model for regenerative medicine. For experimental autologous cell therapy based on induced pluripotent (iPS) cells in the marmoset, cells must be able to undergo robust and reliable directed differentiation that will not require customization for each specific iPS cell clone. When marmoset iPS cells were aggregated in a hanging drop format for 3. days, followed by exposure to dual SMAD inhibitors and retinoic acid in monolayer culture for 3. days, we found substantial variability in the response of different iPS cell clones. However, when clones were pretreated with 0.05-2% dimethyl sulfoxide (DMSO) for 24. hours, all clones showed a very similar maximal response to the directed differentiation scheme. Peak responses were observed at 0.5% DMSO in two clones and at 1% DMSO in a third clone. When patterns of gene expression were examined by microarray analysis, hierarchical clustering showed very similar responses in all 3 clones when they were pretreated with optimal DMSO concentrations. The change in phenotype following exposure to DMSO and the 6. day hanging drop/monolayer treatment was confirmed by immunocytochemistry. Analysis of DNA content in DMSO-exposed cells indicated that it is unlikely that DMSO acts by causing cells to exit from the cell cycle. This approach should be generally valuable in the directed neural differentiation of pluripotent cells for experimental cell therapy.

Original languageEnglish (US)
Pages (from-to)141-150
Number of pages10
JournalStem Cell Research
Volume15
Issue number1
DOIs
StatePublished - Jul 1 2015

Fingerprint

Induced Pluripotent Stem Cells
Callithrix
Dimethyl Sulfoxide
Clone Cells
Cell- and Tissue-Based Therapy
Investigational Therapies
Regenerative Medicine
Microarray Analysis
Tretinoin
Primates
Cluster Analysis
Cell Differentiation
Cell Cycle
Immunohistochemistry
Phenotype
Gene Expression
DNA

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Medicine(all)

Cite this

Qiu, Z., Mishra, A., Li, M., Farnsworth, S. L., Guerra, B., Lanford, R. E., & Hornsby, P. J. (2015). Marmoset induced pluripotent stem cells: Robust neural differentiation following pretreatment with dimethyl sulfoxide. Stem Cell Research, 15(1), 141-150. https://doi.org/10.1016/j.scr.2015.05.010

Marmoset induced pluripotent stem cells : Robust neural differentiation following pretreatment with dimethyl sulfoxide. / Qiu, Zhifang; Mishra, Anuja; Li, Miao; Farnsworth, Steven L.; Guerra, Bernadette; Lanford, Robert E.; Hornsby, Peter J.

In: Stem Cell Research, Vol. 15, No. 1, 01.07.2015, p. 141-150.

Research output: Contribution to journalArticle

Qiu, Z, Mishra, A, Li, M, Farnsworth, SL, Guerra, B, Lanford, RE & Hornsby, PJ 2015, 'Marmoset induced pluripotent stem cells: Robust neural differentiation following pretreatment with dimethyl sulfoxide', Stem Cell Research, vol. 15, no. 1, pp. 141-150. https://doi.org/10.1016/j.scr.2015.05.010
Qiu, Zhifang ; Mishra, Anuja ; Li, Miao ; Farnsworth, Steven L. ; Guerra, Bernadette ; Lanford, Robert E. ; Hornsby, Peter J. / Marmoset induced pluripotent stem cells : Robust neural differentiation following pretreatment with dimethyl sulfoxide. In: Stem Cell Research. 2015 ; Vol. 15, No. 1. pp. 141-150.
@article{2dae63e3d6dc4d489cca615b07370424,
title = "Marmoset induced pluripotent stem cells: Robust neural differentiation following pretreatment with dimethyl sulfoxide",
abstract = "The marmoset is an important nonhuman primate model for regenerative medicine. For experimental autologous cell therapy based on induced pluripotent (iPS) cells in the marmoset, cells must be able to undergo robust and reliable directed differentiation that will not require customization for each specific iPS cell clone. When marmoset iPS cells were aggregated in a hanging drop format for 3. days, followed by exposure to dual SMAD inhibitors and retinoic acid in monolayer culture for 3. days, we found substantial variability in the response of different iPS cell clones. However, when clones were pretreated with 0.05-2{\%} dimethyl sulfoxide (DMSO) for 24. hours, all clones showed a very similar maximal response to the directed differentiation scheme. Peak responses were observed at 0.5{\%} DMSO in two clones and at 1{\%} DMSO in a third clone. When patterns of gene expression were examined by microarray analysis, hierarchical clustering showed very similar responses in all 3 clones when they were pretreated with optimal DMSO concentrations. The change in phenotype following exposure to DMSO and the 6. day hanging drop/monolayer treatment was confirmed by immunocytochemistry. Analysis of DNA content in DMSO-exposed cells indicated that it is unlikely that DMSO acts by causing cells to exit from the cell cycle. This approach should be generally valuable in the directed neural differentiation of pluripotent cells for experimental cell therapy.",
author = "Zhifang Qiu and Anuja Mishra and Miao Li and Farnsworth, {Steven L.} and Bernadette Guerra and Lanford, {Robert E.} and Hornsby, {Peter J}",
year = "2015",
month = "7",
day = "1",
doi = "10.1016/j.scr.2015.05.010",
language = "English (US)",
volume = "15",
pages = "141--150",
journal = "Stem Cell Research",
issn = "1873-5061",
publisher = "Elsevier",
number = "1",

}

TY - JOUR

T1 - Marmoset induced pluripotent stem cells

T2 - Robust neural differentiation following pretreatment with dimethyl sulfoxide

AU - Qiu, Zhifang

AU - Mishra, Anuja

AU - Li, Miao

AU - Farnsworth, Steven L.

AU - Guerra, Bernadette

AU - Lanford, Robert E.

AU - Hornsby, Peter J

PY - 2015/7/1

Y1 - 2015/7/1

N2 - The marmoset is an important nonhuman primate model for regenerative medicine. For experimental autologous cell therapy based on induced pluripotent (iPS) cells in the marmoset, cells must be able to undergo robust and reliable directed differentiation that will not require customization for each specific iPS cell clone. When marmoset iPS cells were aggregated in a hanging drop format for 3. days, followed by exposure to dual SMAD inhibitors and retinoic acid in monolayer culture for 3. days, we found substantial variability in the response of different iPS cell clones. However, when clones were pretreated with 0.05-2% dimethyl sulfoxide (DMSO) for 24. hours, all clones showed a very similar maximal response to the directed differentiation scheme. Peak responses were observed at 0.5% DMSO in two clones and at 1% DMSO in a third clone. When patterns of gene expression were examined by microarray analysis, hierarchical clustering showed very similar responses in all 3 clones when they were pretreated with optimal DMSO concentrations. The change in phenotype following exposure to DMSO and the 6. day hanging drop/monolayer treatment was confirmed by immunocytochemistry. Analysis of DNA content in DMSO-exposed cells indicated that it is unlikely that DMSO acts by causing cells to exit from the cell cycle. This approach should be generally valuable in the directed neural differentiation of pluripotent cells for experimental cell therapy.

AB - The marmoset is an important nonhuman primate model for regenerative medicine. For experimental autologous cell therapy based on induced pluripotent (iPS) cells in the marmoset, cells must be able to undergo robust and reliable directed differentiation that will not require customization for each specific iPS cell clone. When marmoset iPS cells were aggregated in a hanging drop format for 3. days, followed by exposure to dual SMAD inhibitors and retinoic acid in monolayer culture for 3. days, we found substantial variability in the response of different iPS cell clones. However, when clones were pretreated with 0.05-2% dimethyl sulfoxide (DMSO) for 24. hours, all clones showed a very similar maximal response to the directed differentiation scheme. Peak responses were observed at 0.5% DMSO in two clones and at 1% DMSO in a third clone. When patterns of gene expression were examined by microarray analysis, hierarchical clustering showed very similar responses in all 3 clones when they were pretreated with optimal DMSO concentrations. The change in phenotype following exposure to DMSO and the 6. day hanging drop/monolayer treatment was confirmed by immunocytochemistry. Analysis of DNA content in DMSO-exposed cells indicated that it is unlikely that DMSO acts by causing cells to exit from the cell cycle. This approach should be generally valuable in the directed neural differentiation of pluripotent cells for experimental cell therapy.

UR - http://www.scopus.com/inward/record.url?scp=84930948095&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84930948095&partnerID=8YFLogxK

U2 - 10.1016/j.scr.2015.05.010

DO - 10.1016/j.scr.2015.05.010

M3 - Article

C2 - 26070112

AN - SCOPUS:84930948095

VL - 15

SP - 141

EP - 150

JO - Stem Cell Research

JF - Stem Cell Research

SN - 1873-5061

IS - 1

ER -