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ZILOS-tk Laser | XYClone Laser | CASA | DNA Technology

XYClone Articles, Papers and Posters

A complete searchable database of all papers featuring Hamitlon Thorne CASA and Laser systems may be accessed on the Connotea web site at:
http://www.connotea.org/user/hamiltonthorne

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NEW! January 2007

F0 generation mice fully derived from gene-targeted embryonic stem cells allowing immediate phenotypic analyses.
William T Poueymirou, Wojtek Auerbach, David Frendewey, Joseph F Hickey, Jennifer M Escaravage, Lakeisha Esau, Anthony T Dore´, Sean Stevens, Niels C Adams, Melissa G Dominguez, Nicholas W Gale, George D Yancopoulos, Thomas M DeChiara& David M Valenzuela.

Nature Biotechnology, 2007. Jan. 25(1):91-99.

A useful approach for exploring gene function involves generating mutant mice from genetically modified embryonic stem (ES) cells. Recent advances in genetic engineering of ES cells have shifted the bottleneck in this process to the generation of mice. Conventional injections of ES cells into blastocyst hosts produce F0 generation chimeras that are only partially derived from ES cells, requiring additional breeding to obtain mutant mice that can be phenotyped. The tetraploid complementation approach directly yields mice that are almost entirely derived from ES cells, but it is inefficient, works only with certain hybrid ES cell lines and suffers from nonspecific lethality and abnormalities, complicating phenotypic analyses. Here we show that laserassisted injection of either inbred or hybrid ES cells into eight cell–stage embryos efficiently yields F0 generation mice that are fully ES cell–derived and healthy, exhibit 100% germline transmission and allow immediate phenotypic analysis, greatly accelerating gene function assignment.

NEW! January 2007

Human Reproduction, 2007. Jan. 22(1):52–62.

Developmental competence of human in vitro aged oocytes as host cells for nuclear transfer.
V.J.Hall, D.Compton, P.Stojkovic, M.Nesbitt, M.Herbert, A.Murdoch and M.Stojkovic

BACKGROUND: Improving human nuclear transfer (NT) efficiencies is paramount for the development of patient-specific stem cell lines, although the opportunities remain limited owing to difficulties in obtaining fresh mature oocytes. METHODS: Therefore, the developmental competence of aged, failed-to-fertilize human oocytes as an alternate cytoplasmic source for NT was assessed and compared with use of fresh, ovulation-induced oocytes. To further characterize the developmental potential of aged oocytes, parthenogenetic activation, immunocytochemical analysis of essential microtubule proteins involved in meiotic and mitotic division, and RT–PCR in single oocytes (n = 6) was performed to determine expression of oocyte-specific genes [oocyte-specific histone 1 (H1FOO), growth differentiation factor 9 (GDF9), bone morphogenetic protein 15 (BMP15), zygote arrest 1 (ZAR1)] and microtubule markers [nuclear mitotic arrest (NuMA), minus-end directed motor protein HSET and the microtubule kinesin motor protein EG5]. RESULTS: For NT, enucleation and fusion rates of aged oocytes were significantly lower compared with fresh oocytes (P < 0.05). Cleavage rates and subsequent development were poor. In addition, parthenote cleavage was low. Immunocytochemical analysis revealed that many oocytes displayed aberrant expression of NuMA and EG5, had disrupted meiotic spindles and tetrapolar spindles. One of the six oocytes misexpressed GDF9, BMP15 and ZAR1. Two oocytes expressed EG5 messenger RNA (mRNA), and HSET and NuMA were not detectable. RT-PCR of mRNA for oocyte specific genes and microtubule markers in single aged oocytes. CONCLUSIONS: Thus, aneuploidy and spindle defects may contribute to poor parthenogenetic development and developmental outcomes following NT.

NEW! January 2007

Reproduction, Fertility and Development 19(1) 225 - 226 (abstracts from IETS 2007)

217 A NEW APPROACH TO CRYOPRESERVATION OF LARGE EQUINE EMBRYOS BY VITRIFICATION AFTER BLASTOCOEL MICROMANIPULATION
J. Scherzer, R. A. Fayrer-Hosken, L. Ray and G. Heusner
(Download PDF of abstract)

2006

Laser-assisted blastocyst dissection and subsequent cultivation of embryonic stem cells in a serum/cell free culture system: applications and preliminary results in a murine model. (Download PDF of Full Paper)
Noriko Tanaka1, Takumi Takeuchi1, Queenie V Neri1, Eric Scott Sills2 and Gianpiero D Palermo1
1Center for Reproductive Medicine and Infertility, Weill Medical College of Cornell University, New York, NY 10021, USA
2Department of Obstetrics, Gynecology and Reproductive Research, Murphy Medical Center, Murphy, NC, USA
Journal of Translational Medicine 2006, 4 : 20     doi:10.1186/1479-5876-4-20
http://www.translational-medicine.com/content/4/1/20

Effects of Caffeine on the In Vitro Development of Bovine Nuclear Transfer Embryos. Maalouf WE, Lee J-H, Campbell KHS. Animal Development and Biotechnology Group, School of Biosciences, University of Nottingham. Presented at IETS 2006. (View Poster PDF)

2005

"Laser zona drilling does not induce hsp70i transcription in blastomeres of eight-cell mouse embryos." Cristina Hartshorn, Ph.D., Aleksandra Anshelevich, B.S.,
Lawrence J. Wangh, Ph.D. Note: This study used the ZILOS-tk, sister product to the newer XYClone system. The XYClone is now recommended for all animal work.

To assess whether zona drilling with a 1,480-nm laser induces heat shock in eight-cell embryos, we measured hsp70i RNA levels in sets of single blastomeres isolated after laser treatment of mouse embryos that had or had not been heated at 43°C. Unlike heating, laser zona drilling did not stimulate hsp70i expression, even in the blastomeres closest to the laser beam, corroborating the safety of this procedure for assisted reproduction. (Fertil Steril 2005;84:1547–50. ©2005 by American Society for Reproductive Medicine.) (View Full Article)

A New Method to Produce Mice Fully Derived from Embryonic Stem Cells. William T. Poueymirou, Wojtek Auerbach, Joseph Hickey, Jennifer Escaravage, David Frendewey, Niels C. Adams, George D. Yancopoulos, Thomad D. DeChiara, and David M. Valenzuela. Regerneron Pharmaceuticals, Inc. 777 Old Saw Mill River Road., tarrytown, NY, USA. Presented at the 2005 Transgenic Technology Meeting, Barcelona, Spain, September 11-13, 2005.(View the Full Poster PDF)

A Feeder Cell- and Serum-Free System to Harvest Embryonic Stem Cells From Laser Dissected Blastocysts. T. Takeuchi, N. Tanaka, Q.V. Neri, Z. Rosenwaks, G.D. Palermo. Cornell University, New York, NY. Abstract from American Society of Reproductive Medicine, September 2005 (View Abstract PDF)

XYClone™ Laser System: A Novel Technique for Mouse Intracytoplasmic Sperm Injection (ICSI). William A. Richie, Embryologist, Division of Gene Expression and Development, Roslin Institute, Roslin Midlothian UK EH25 9PS and K. Bradley, Hamilton Thorne Biosciences, Beverly, MA 01915 USA (View Poster PDF)