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Dev Dyn
2009 Oct 01;23810:2614-21. doi: 10.1002/dvdy.22080.
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Enucleation of feeder cells and egg cells with psoralens.
McGarry TJ
,
Bonaguidi M
,
Lyass L
,
Kessler JA
,
Bodily JM
,
Doglio L
.
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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.
Cimino,
Psoralens as photoactive probes of nucleic acid structure and function: organic chemistry, photochemistry, and biochemistry.
1985, Pubmed
Cimino,
Psoralens as photoactive probes of nucleic acid structure and function: organic chemistry, photochemistry, and biochemistry.
1985,
Pubmed
Cowan,
Nuclear reprogramming of somatic cells after fusion with human embryonic stem cells.
2005,
Pubmed
Darzynkiewicz,
DNA content measurement for DNA ploidy and cell cycle analysis.
2001,
Pubmed
Fulka,
Noninvasive chemical enucleation of mouse oocytes.
1993,
Pubmed
Gerhart,
Cell cycle dynamics of an M-phase-specific cytoplasmic factor in Xenopus laevis oocytes and eggs.
1984,
Pubmed
,
Xenbase
Gurdon,
Nuclear transplantation in Xenopus.
1991,
Pubmed
,
Xenbase
Kane,
The zebrafish midblastula transition.
1993,
Pubmed
Kawakami,
Effect of demecolcine and nocodazole on the efficiency of chemically assisted removal of chromosomes and the developmental potential of nuclear transferred porcine oocytes.
2003,
Pubmed
Kroll,
Transgenic X. laevis embryos from eggs transplanted with nuclei of transfected cultured cells.
1994,
Pubmed
,
Xenbase
Li,
Chemically assisted handmade enucleation of porcine oocytes.
2006,
Pubmed
Li,
Review of enucleation methods and procedures used in animal cloning: state of the art.
2004,
Pubmed
Liu,
A reliable, noninvasive technique for spindle imaging and enucleation of mammalian oocytes.
2000,
Pubmed
Liu,
Chk1 is an essential kinase that is regulated by Atr and required for the G(2)/M DNA damage checkpoint.
2000,
Pubmed
Marsh-Armstrong,
Germ-line transmission of transgenes in Xenopus laevis.
1999,
Pubmed
,
Xenbase
Murray,
Cell cycle extracts.
1991,
Pubmed
O'Farrell,
Embryonic cleavage cycles: how is a mouse like a fly?
2004,
Pubmed
Ozbun,
Infectious human papillomavirus type 31b: purification and infection of an immortalized human keratinocyte cell line.
2002,
Pubmed
Pichierri,
The DNA crosslink-induced S-phase checkpoint depends on ATR-CHK1 and ATR-NBS1-FANCD2 pathways.
2004,
Pubmed
Quinn,
Review of media used in ART laboratories.
2000,
Pubmed
Raff,
Nuclear and cytoplasmic mitotic cycles continue in Drosophila embryos in which DNA synthesis is inhibited with aphidicolin.
1988,
Pubmed
Smith,
Membrane and intracellular effects of ultraviolet irradiation with Hoechst 33342 on bovine secondary oocytes matured in vitro.
1993,
Pubmed
Stern,
Psoralen and ultraviolet a light therapy for psoriasis.
2007,
Pubmed
Takai,
Aberrant cell cycle checkpoint function and early embryonic death in Chk1(-/-) mice.
2000,
Pubmed
Tatham,
Enucleation by centrifugation of in vitro-matured bovine oocytes for use in nuclear transfer.
1995,
Pubmed
Thomson,
Embryonic stem cell lines derived from human blastocysts.
1998,
Pubmed
Tymowska,
Karyotype analysis of Xenopus muelleri (Peters) and Xenopus laevis (Daudin), Pipidae.
1972,
Pubmed
,
Xenbase
Vajta,
Highly efficient and reliable chemically assisted enucleation method for handmade cloning in cattle.
2005,
Pubmed
Villa-Diaz,
Analysis of the factors that limit the ability of feeder cells to maintain the undifferentiated state of human embryonic stem cells.
2009,
Pubmed
Wang,
Sucrose pretreatment for enucleation: an efficient and non-damage method for removing the spindle of the mouse MII oocyte.
2001,
Pubmed