IVF recovery of mutant mouse lines using sperm cryopreserved with MTG In cryovials

Ming Wen Li, Jadine M. Vallelunga, Kristy L. Kinchen, Karina L. Rink, Jasmin Zarrabi, Armen O. Shamamian, Kevin C K Lloyd

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

BACKGROUND: Modification of cryoprotective medium (CPM) R18S3 (18% raffinose and 3% skim milk) by addition of monothioglycerol (MTG) or L-glutamine (Glu) has been shown to improve in vitro fertilization (IVF) using mouse sperm cryopreserved in cryostraws. However, whether these CPMs can be applied effectively to sperm cryopreserved in cryovials is unknown. OBJECTIVE: The study was to determine the comparative effectiveness of using R18S3, R18S3+Glu (lOOmM and 87 mM), or R18S3+MTG (477 (aM) to cryopreserve various sample volumes of mouse sperm in cryovials and cryostraws. METHODS: This study compared the effects of different CPMs on motility of fresh and frozen-thawed C57BL/6J sperm and on IVF rate of C57BL/6J sperm cryopreserved in different CPMs and containers with different volumes, and then used technologies developed to cryopreserve and recover sperm of knockout mouse lines on inbred C57BL/6 backgrounds. RESULTS: Glutamine at 100 mM inhibited, but MTG at 477 fiM protected, fresh sperm motility significantly (P<0.05). Sperm cryopreserved in R18S3+MTG had significantly better (P<0.05) post-thaw progressive motility and IVF rate than when cryopreserved in R18S3 alone, R18S3+Glu (100 mM), or RSGlu87 (15.7% raffinose, 2.6% skim milk, and 87 mM L-glutamine). There was no significant difference in IVF rates among sperm cryopreserved with R18S3+MTG in cryovials or in cryostraws (P>0.05). Sperm from 63 knockout mouse lines on C57BL/6 backgrounds cryopreserved using R18S3+MTG in cryovials were all recovered successfully to genotypically-confirmed offspring. CONCLUSION: Mouse sperm on C56BL/6 backgrounds can be successfully cryopreserved in cryovials using R18S3+MTG.

Original languageEnglish (US)
Pages (from-to)145-153
Number of pages9
JournalCryo-Letters
Volume35
Issue number2
StatePublished - 2014

Fingerprint

in vitro fertilization
Fertilization in Vitro
Spermatozoa
spermatozoa
mutants
mice
Glutamine
glutamine
Knockout Mice
Raffinose
thioglycerol
Sperm Motility
raffinose
skim milk
sperm motility
containers
Milk
Technology

Keywords

  • Cryopreservation
  • Cryostraw
  • Cryovial
  • IVF
  • Mouse
  • Sperm

ASJC Scopus subject areas

  • Agronomy and Crop Science
  • Anatomy

Cite this

Li, M. W., Vallelunga, J. M., Kinchen, K. L., Rink, K. L., Zarrabi, J., Shamamian, A. O., & Lloyd, K. C. K. (2014). IVF recovery of mutant mouse lines using sperm cryopreserved with MTG In cryovials. Cryo-Letters, 35(2), 145-153.

IVF recovery of mutant mouse lines using sperm cryopreserved with MTG In cryovials. / Li, Ming Wen; Vallelunga, Jadine M.; Kinchen, Kristy L.; Rink, Karina L.; Zarrabi, Jasmin; Shamamian, Armen O.; Lloyd, Kevin C K.

In: Cryo-Letters, Vol. 35, No. 2, 2014, p. 145-153.

Research output: Contribution to journalArticle

Li, MW, Vallelunga, JM, Kinchen, KL, Rink, KL, Zarrabi, J, Shamamian, AO & Lloyd, KCK 2014, 'IVF recovery of mutant mouse lines using sperm cryopreserved with MTG In cryovials', Cryo-Letters, vol. 35, no. 2, pp. 145-153.
Li MW, Vallelunga JM, Kinchen KL, Rink KL, Zarrabi J, Shamamian AO et al. IVF recovery of mutant mouse lines using sperm cryopreserved with MTG In cryovials. Cryo-Letters. 2014;35(2):145-153.
Li, Ming Wen ; Vallelunga, Jadine M. ; Kinchen, Kristy L. ; Rink, Karina L. ; Zarrabi, Jasmin ; Shamamian, Armen O. ; Lloyd, Kevin C K. / IVF recovery of mutant mouse lines using sperm cryopreserved with MTG In cryovials. In: Cryo-Letters. 2014 ; Vol. 35, No. 2. pp. 145-153.
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abstract = "BACKGROUND: Modification of cryoprotective medium (CPM) R18S3 (18{\%} raffinose and 3{\%} skim milk) by addition of monothioglycerol (MTG) or L-glutamine (Glu) has been shown to improve in vitro fertilization (IVF) using mouse sperm cryopreserved in cryostraws. However, whether these CPMs can be applied effectively to sperm cryopreserved in cryovials is unknown. OBJECTIVE: The study was to determine the comparative effectiveness of using R18S3, R18S3+Glu (lOOmM and 87 mM), or R18S3+MTG (477 (aM) to cryopreserve various sample volumes of mouse sperm in cryovials and cryostraws. METHODS: This study compared the effects of different CPMs on motility of fresh and frozen-thawed C57BL/6J sperm and on IVF rate of C57BL/6J sperm cryopreserved in different CPMs and containers with different volumes, and then used technologies developed to cryopreserve and recover sperm of knockout mouse lines on inbred C57BL/6 backgrounds. RESULTS: Glutamine at 100 mM inhibited, but MTG at 477 fiM protected, fresh sperm motility significantly (P<0.05). Sperm cryopreserved in R18S3+MTG had significantly better (P<0.05) post-thaw progressive motility and IVF rate than when cryopreserved in R18S3 alone, R18S3+Glu (100 mM), or RSGlu87 (15.7{\%} raffinose, 2.6{\%} skim milk, and 87 mM L-glutamine). There was no significant difference in IVF rates among sperm cryopreserved with R18S3+MTG in cryovials or in cryostraws (P>0.05). Sperm from 63 knockout mouse lines on C57BL/6 backgrounds cryopreserved using R18S3+MTG in cryovials were all recovered successfully to genotypically-confirmed offspring. CONCLUSION: Mouse sperm on C56BL/6 backgrounds can be successfully cryopreserved in cryovials using R18S3+MTG.",
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AU - Li, Ming Wen

AU - Vallelunga, Jadine M.

AU - Kinchen, Kristy L.

AU - Rink, Karina L.

AU - Zarrabi, Jasmin

AU - Shamamian, Armen O.

AU - Lloyd, Kevin C K

PY - 2014

Y1 - 2014

N2 - BACKGROUND: Modification of cryoprotective medium (CPM) R18S3 (18% raffinose and 3% skim milk) by addition of monothioglycerol (MTG) or L-glutamine (Glu) has been shown to improve in vitro fertilization (IVF) using mouse sperm cryopreserved in cryostraws. However, whether these CPMs can be applied effectively to sperm cryopreserved in cryovials is unknown. OBJECTIVE: The study was to determine the comparative effectiveness of using R18S3, R18S3+Glu (lOOmM and 87 mM), or R18S3+MTG (477 (aM) to cryopreserve various sample volumes of mouse sperm in cryovials and cryostraws. METHODS: This study compared the effects of different CPMs on motility of fresh and frozen-thawed C57BL/6J sperm and on IVF rate of C57BL/6J sperm cryopreserved in different CPMs and containers with different volumes, and then used technologies developed to cryopreserve and recover sperm of knockout mouse lines on inbred C57BL/6 backgrounds. RESULTS: Glutamine at 100 mM inhibited, but MTG at 477 fiM protected, fresh sperm motility significantly (P<0.05). Sperm cryopreserved in R18S3+MTG had significantly better (P<0.05) post-thaw progressive motility and IVF rate than when cryopreserved in R18S3 alone, R18S3+Glu (100 mM), or RSGlu87 (15.7% raffinose, 2.6% skim milk, and 87 mM L-glutamine). There was no significant difference in IVF rates among sperm cryopreserved with R18S3+MTG in cryovials or in cryostraws (P>0.05). Sperm from 63 knockout mouse lines on C57BL/6 backgrounds cryopreserved using R18S3+MTG in cryovials were all recovered successfully to genotypically-confirmed offspring. CONCLUSION: Mouse sperm on C56BL/6 backgrounds can be successfully cryopreserved in cryovials using R18S3+MTG.

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KW - Cryostraw

KW - Cryovial

KW - IVF

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