Enucleation of feeder cells and egg cells with psoralens

Thomas J. McGarry, Michael Bonaguidi, Ljuba Lyass, John A. Kessler, Jason M. Bodily, Lynn Doglio

Research output: Contribution to journalArticle

Abstract

The cell nucleus must be inactivated or destroyed in order to generate feeder layers for cultured cells or to prepare recipient egg cells for nuclear transfer. Existing enucleation techniques are either cumbersome or employ toxic chemicals. Here we report a new method to enucleate cells by treatment with a psoralen and long-wave ultraviolet light. The technique is >90% efficient and causes little cytoplasmic damage to the treated cell. We have used psoralen treatment to enucleate a wide variety of cells, including eggs, sperm, HeLa cells, and fibroblasts. Colonies of human embryonic stem cells (hESCs) and human keratinocyte precursors grown on psoralen-treated feeders are indistinguishable from those grown on gamma-irradiated or mitomycin C-treated cells. Psoralen enucleation provides a rapid, simple, and non-toxic method to generate feeder cells. The technique is also useful for nuclear transfer studies in species with large eggs whose cleavage divisions are not regulated by cell-cycle checkpoints.

Original languageEnglish (US)
Pages (from-to)2614-2621
Number of pages8
JournalDevelopmental Dynamics
Volume238
Issue number10
DOIs
StatePublished - Oct 2009
Externally publishedYes

Fingerprint

Feeder Cells
Ficusin
Ovum
Eggs
Poisons
Mitomycin
Ultraviolet Rays
Cell Cycle Checkpoints
Cell Nucleus
Keratinocytes
HeLa Cells
Spermatozoa
Cultured Cells
Fibroblasts
Furocoumarins

Keywords

  • Cell culture
  • Enucleation
  • Feeder cells
  • Nuclear transplantation
  • Psoralen
  • Ultraviolet light

ASJC Scopus subject areas

  • Developmental Biology

Cite this

McGarry, T. J., Bonaguidi, M., Lyass, L., Kessler, J. A., Bodily, J. M., & Doglio, L. (2009). Enucleation of feeder cells and egg cells with psoralens. Developmental Dynamics, 238(10), 2614-2621. https://doi.org/10.1002/dvdy.22080

Enucleation of feeder cells and egg cells with psoralens. / McGarry, Thomas J.; Bonaguidi, Michael; Lyass, Ljuba; Kessler, John A.; Bodily, Jason M.; Doglio, Lynn.

In: Developmental Dynamics, Vol. 238, No. 10, 10.2009, p. 2614-2621.

Research output: Contribution to journalArticle

McGarry, TJ, Bonaguidi, M, Lyass, L, Kessler, JA, Bodily, JM & Doglio, L 2009, 'Enucleation of feeder cells and egg cells with psoralens', Developmental Dynamics, vol. 238, no. 10, pp. 2614-2621. https://doi.org/10.1002/dvdy.22080
McGarry TJ, Bonaguidi M, Lyass L, Kessler JA, Bodily JM, Doglio L. Enucleation of feeder cells and egg cells with psoralens. Developmental Dynamics. 2009 Oct;238(10):2614-2621. https://doi.org/10.1002/dvdy.22080
McGarry, Thomas J. ; Bonaguidi, Michael ; Lyass, Ljuba ; Kessler, John A. ; Bodily, Jason M. ; Doglio, Lynn. / Enucleation of feeder cells and egg cells with psoralens. In: Developmental Dynamics. 2009 ; Vol. 238, No. 10. pp. 2614-2621.
@article{4088c7a830c541128ae14712bda819d5,
title = "Enucleation of feeder cells and egg cells with psoralens",
abstract = "The cell nucleus must be inactivated or destroyed in order to generate feeder layers for cultured cells or to prepare recipient egg cells for nuclear transfer. Existing enucleation techniques are either cumbersome or employ toxic chemicals. Here we report a new method to enucleate cells by treatment with a psoralen and long-wave ultraviolet light. The technique is >90{\%} efficient and causes little cytoplasmic damage to the treated cell. We have used psoralen treatment to enucleate a wide variety of cells, including eggs, sperm, HeLa cells, and fibroblasts. Colonies of human embryonic stem cells (hESCs) and human keratinocyte precursors grown on psoralen-treated feeders are indistinguishable from those grown on gamma-irradiated or mitomycin C-treated cells. Psoralen enucleation provides a rapid, simple, and non-toxic method to generate feeder cells. The technique is also useful for nuclear transfer studies in species with large eggs whose cleavage divisions are not regulated by cell-cycle checkpoints.",
keywords = "Cell culture, Enucleation, Feeder cells, Nuclear transplantation, Psoralen, Ultraviolet light",
author = "McGarry, {Thomas J.} and Michael Bonaguidi and Ljuba Lyass and Kessler, {John A.} and Bodily, {Jason M.} and Lynn Doglio",
year = "2009",
month = "10",
doi = "10.1002/dvdy.22080",
language = "English (US)",
volume = "238",
pages = "2614--2621",
journal = "Developmental Dynamics",
issn = "1058-8388",
publisher = "Wiley-Liss Inc.",
number = "10",

}

TY - JOUR

T1 - Enucleation of feeder cells and egg cells with psoralens

AU - McGarry, Thomas J.

AU - Bonaguidi, Michael

AU - Lyass, Ljuba

AU - Kessler, John A.

AU - Bodily, Jason M.

AU - Doglio, Lynn

PY - 2009/10

Y1 - 2009/10

N2 - The cell nucleus must be inactivated or destroyed in order to generate feeder layers for cultured cells or to prepare recipient egg cells for nuclear transfer. Existing enucleation techniques are either cumbersome or employ toxic chemicals. Here we report a new method to enucleate cells by treatment with a psoralen and long-wave ultraviolet light. The technique is >90% efficient and causes little cytoplasmic damage to the treated cell. We have used psoralen treatment to enucleate a wide variety of cells, including eggs, sperm, HeLa cells, and fibroblasts. Colonies of human embryonic stem cells (hESCs) and human keratinocyte precursors grown on psoralen-treated feeders are indistinguishable from those grown on gamma-irradiated or mitomycin C-treated cells. Psoralen enucleation provides a rapid, simple, and non-toxic method to generate feeder cells. The technique is also useful for nuclear transfer studies in species with large eggs whose cleavage divisions are not regulated by cell-cycle checkpoints.

AB - The cell nucleus must be inactivated or destroyed in order to generate feeder layers for cultured cells or to prepare recipient egg cells for nuclear transfer. Existing enucleation techniques are either cumbersome or employ toxic chemicals. Here we report a new method to enucleate cells by treatment with a psoralen and long-wave ultraviolet light. The technique is >90% efficient and causes little cytoplasmic damage to the treated cell. We have used psoralen treatment to enucleate a wide variety of cells, including eggs, sperm, HeLa cells, and fibroblasts. Colonies of human embryonic stem cells (hESCs) and human keratinocyte precursors grown on psoralen-treated feeders are indistinguishable from those grown on gamma-irradiated or mitomycin C-treated cells. Psoralen enucleation provides a rapid, simple, and non-toxic method to generate feeder cells. The technique is also useful for nuclear transfer studies in species with large eggs whose cleavage divisions are not regulated by cell-cycle checkpoints.

KW - Cell culture

KW - Enucleation

KW - Feeder cells

KW - Nuclear transplantation

KW - Psoralen

KW - Ultraviolet light

UR - http://www.scopus.com/inward/record.url?scp=70349329564&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=70349329564&partnerID=8YFLogxK

U2 - 10.1002/dvdy.22080

DO - 10.1002/dvdy.22080

M3 - Article

C2 - 19705441

AN - SCOPUS:70349329564

VL - 238

SP - 2614

EP - 2621

JO - Developmental Dynamics

JF - Developmental Dynamics

SN - 1058-8388

IS - 10

ER -