Cryopreserved morulae can be used to efficiently generate germline-transmitting chimeras by blastocyst injection

Janice V. Parker-Thornburg, Jennifer L. Alana, Chad N. Smith, Michelle Detry, Marta L. Rojas, Kedryn K. Baskin

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

The production of chimeric mice is a complex process, requiring the careful coordination of tissue culture cell growth, production of a large number (30-75) of competent blastocysts and the availability of appropriately timed pseudo pregnant female mice. Failure at any of these steps can impinge upon the rapid production of chimeras. One potential improvement for the efficient generation of chimeric mice would be the utilization of cryopreserved embryos suitable for injection. C57Bl/6 morulae were frozen using a standard 2-step protocol with ethylene glycol as the cryopreservation agent. We determined that cryopreserved morulae could thaw, culture to blastocyst stage in KSOM media and survive injection at rates equivalent to control embryos. Cryopreserved morulae were also equivalent to controls at all later stages in the process of production of chimeric mice, including birth rate, percentage chimerism of resulting animals and ability to produce germline progeny. Hence, cryopreservation of morulae for blastocyst injection is a suitable option to enhance the efficiency of chimeric mouse generation.

Original languageEnglish (US)
Pages (from-to)685-690
Number of pages6
JournalTransgenic Research
Volume14
Issue number5
DOIs
StatePublished - Oct 1 2005

Fingerprint

Morula
chimerism
morula
Blastocyst
blastocyst
germ cells
injection
Injections
mice
Cryopreservation
cryopreservation
embryo (animal)
Embryonic Structures
Chimerism
birth rate
Ethylene Glycol
Birth Rate
ethylene glycol
tissue culture
cell growth

Keywords

  • Blastocyst injection
  • Cryopreservation
  • ES cells
  • Embryo freezing
  • Germline transmission

ASJC Scopus subject areas

  • Biotechnology
  • Animal Science and Zoology
  • Agronomy and Crop Science
  • Genetics

Cite this

Cryopreserved morulae can be used to efficiently generate germline-transmitting chimeras by blastocyst injection. / Parker-Thornburg, Janice V.; Alana, Jennifer L.; Smith, Chad N.; Detry, Michelle; Rojas, Marta L.; Baskin, Kedryn K.

In: Transgenic Research, Vol. 14, No. 5, 01.10.2005, p. 685-690.

Research output: Contribution to journalArticle

Parker-Thornburg, Janice V. ; Alana, Jennifer L. ; Smith, Chad N. ; Detry, Michelle ; Rojas, Marta L. ; Baskin, Kedryn K. / Cryopreserved morulae can be used to efficiently generate germline-transmitting chimeras by blastocyst injection. In: Transgenic Research. 2005 ; Vol. 14, No. 5. pp. 685-690.
@article{3cd6fd57a7bb44318f11181631ac1f70,
title = "Cryopreserved morulae can be used to efficiently generate germline-transmitting chimeras by blastocyst injection",
abstract = "The production of chimeric mice is a complex process, requiring the careful coordination of tissue culture cell growth, production of a large number (30-75) of competent blastocysts and the availability of appropriately timed pseudo pregnant female mice. Failure at any of these steps can impinge upon the rapid production of chimeras. One potential improvement for the efficient generation of chimeric mice would be the utilization of cryopreserved embryos suitable for injection. C57Bl/6 morulae were frozen using a standard 2-step protocol with ethylene glycol as the cryopreservation agent. We determined that cryopreserved morulae could thaw, culture to blastocyst stage in KSOM media and survive injection at rates equivalent to control embryos. Cryopreserved morulae were also equivalent to controls at all later stages in the process of production of chimeric mice, including birth rate, percentage chimerism of resulting animals and ability to produce germline progeny. Hence, cryopreservation of morulae for blastocyst injection is a suitable option to enhance the efficiency of chimeric mouse generation.",
keywords = "Blastocyst injection, Cryopreservation, ES cells, Embryo freezing, Germline transmission",
author = "Parker-Thornburg, {Janice V.} and Alana, {Jennifer L.} and Smith, {Chad N.} and Michelle Detry and Rojas, {Marta L.} and Baskin, {Kedryn K.}",
year = "2005",
month = "10",
day = "1",
doi = "10.1007/s11248-005-7022-6",
language = "English (US)",
volume = "14",
pages = "685--690",
journal = "Transgenic Research",
issn = "0962-8819",
publisher = "Springer Netherlands",
number = "5",

}

TY - JOUR

T1 - Cryopreserved morulae can be used to efficiently generate germline-transmitting chimeras by blastocyst injection

AU - Parker-Thornburg, Janice V.

AU - Alana, Jennifer L.

AU - Smith, Chad N.

AU - Detry, Michelle

AU - Rojas, Marta L.

AU - Baskin, Kedryn K.

PY - 2005/10/1

Y1 - 2005/10/1

N2 - The production of chimeric mice is a complex process, requiring the careful coordination of tissue culture cell growth, production of a large number (30-75) of competent blastocysts and the availability of appropriately timed pseudo pregnant female mice. Failure at any of these steps can impinge upon the rapid production of chimeras. One potential improvement for the efficient generation of chimeric mice would be the utilization of cryopreserved embryos suitable for injection. C57Bl/6 morulae were frozen using a standard 2-step protocol with ethylene glycol as the cryopreservation agent. We determined that cryopreserved morulae could thaw, culture to blastocyst stage in KSOM media and survive injection at rates equivalent to control embryos. Cryopreserved morulae were also equivalent to controls at all later stages in the process of production of chimeric mice, including birth rate, percentage chimerism of resulting animals and ability to produce germline progeny. Hence, cryopreservation of morulae for blastocyst injection is a suitable option to enhance the efficiency of chimeric mouse generation.

AB - The production of chimeric mice is a complex process, requiring the careful coordination of tissue culture cell growth, production of a large number (30-75) of competent blastocysts and the availability of appropriately timed pseudo pregnant female mice. Failure at any of these steps can impinge upon the rapid production of chimeras. One potential improvement for the efficient generation of chimeric mice would be the utilization of cryopreserved embryos suitable for injection. C57Bl/6 morulae were frozen using a standard 2-step protocol with ethylene glycol as the cryopreservation agent. We determined that cryopreserved morulae could thaw, culture to blastocyst stage in KSOM media and survive injection at rates equivalent to control embryos. Cryopreserved morulae were also equivalent to controls at all later stages in the process of production of chimeric mice, including birth rate, percentage chimerism of resulting animals and ability to produce germline progeny. Hence, cryopreservation of morulae for blastocyst injection is a suitable option to enhance the efficiency of chimeric mouse generation.

KW - Blastocyst injection

KW - Cryopreservation

KW - ES cells

KW - Embryo freezing

KW - Germline transmission

UR - http://www.scopus.com/inward/record.url?scp=27144489637&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=27144489637&partnerID=8YFLogxK

U2 - 10.1007/s11248-005-7022-6

DO - 10.1007/s11248-005-7022-6

M3 - Article

C2 - 16245159

AN - SCOPUS:27144489637

VL - 14

SP - 685

EP - 690

JO - Transgenic Research

JF - Transgenic Research

SN - 0962-8819

IS - 5

ER -