Open AccessCryobiology of Rat Embryos I: Determination of Zygote Membrane Permeability Coefficients for Water and Cryoprotectants, Their Activation Energies, and the Development of Improved Cryopreservation Methods1
Author(s) -
Reinhold Thomas Pfaff,
Yüksel Ağca,
Jun Li,
Erik J. Woods,
Augustine T. Peter,
John K. Critser
Publication year - 2000
Publication title -
biology of reproduction
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.366
H-Index - 180
eISSN - 1529-7268
pISSN - 0006-3363
DOI - 10.1095/biolreprod63.5.1294
Subject(s) - cryobiology , cryoprotectant , cryopreservation , dimethyl sulfoxide , zygote , vitrification , biology , embryo , ethylene glycol , andrology , microbiology and biotechnology , embryogenesis , chemistry , medicine , organic chemistry
New rat models are being developed at an exponential rate, making improved methods to cryopreserve rat embryos extremely important. However, cryopreservation of rat embryos has proven to be difficult and expensive. In this study, a series of experiments was performed to characterize the fundamental cryobiology of rat fertilized 1-cell embryos (zygotes) and to investigate the effects of different cryoprotective agents (CPAs) and two different plunging temperatures (T(p)) on post-thaw survival of embryos from three genetic backgrounds. In the initial experiments, information on the fundamental cryobiology of rat zygotes was determined, including 1) the hydraulic conductivity in the presence of CPAs (L(p)), 2) the cryoprotectant permeability (P(CPA)), 3) the reflection coefficient (sigma), and 4) the activation energies for these parameters. P(CPA) values were determined for the CPAs, ethylene glycol (EG), dimethyl sulfoxide (DMSO), and propylene glycol (PG). Using this information, a cryopreservation method was developed and the cryosurvival and fetal development of Sprague-Dawley zygotes cryopreserved in either EG, DMSO, or PG and plunged at either -30 or -80 degrees C, were assessed. The highest fetal developmental rates were obtained using a T(p) of -30 degrees C and EG (61.2% +/- 2.4%), which was not different (P > 0.05) from nonfrozen control zygotes (54.6% +/- 3.0%).