Effect of Dextrans on Cryopreservation of Goat Cauda Epididymal Spermatozoa using a Chemically Defined Medium
IR@IICB: CSIR-Indian Institute of Chemical Biology, Kolkata
View Archive Info| Field | Value | |
| Title |
Effect of Dextrans on Cryopreservation of Goat Cauda Epididymal
Spermatozoa using a Chemically Defined Medium
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| Creator |
Kundu, C N
Chakrabarty, J Dutta, P Bhattacharyya, D Ghosh, A Majumder, Gopal Chandra |
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| Subject |
Cell Biology & Physiology
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| Description |
Experiments were carried out to investigate the cryoprotecting
potential of dextrans (ranging from 10 to
2000 kDa) using a synthetic model system developed
recently in this laboratory. Goat spermatozoa from the
cauda epididymidis were extracted in a chemically defined
medium (modified Ringer’s solution) and assayed for
motility using a phase-contrast microscope. The sperm
cells were subjected to a freezing protocol in a computercontrolled
biofreezer (cooling at 0.258C min–1 to 58C; 58C
min–1 to –208C; 208C min–1 to –1008C) and plunged into
liquid nitrogen. The frozen cells were thawed rapidly
at 378C in a thermostatic waterbath. In the absence of
dextran, all the spermatozoa lost their motility. The cryoprotecting
efficacy of each dextran was found to be
biphasic in nature. Initially, as the concentration of
dextran was increased, the recovery of sperm motility also
increased and reached an optimum value; however, with
further increases in dextran concentration, the recovery of
motility decreased sharply. Of all the sugar polymers
tested, 10 kDa dextran showed the highest cryoprotecting
efficacy, whereas the 2000 kDa sugar polymer was the least active. Dextrans of 10, 40, 73, 173, 252, 500 and
2000 kDa offered maximum cryorecovery of forward
motility to the extent of approximately 23%, 21%, 19%,
18%, 16%, 15% and 8%, respectively. Optimum concentrations
of these dextrans for cryoprotection of sperm
motility were 8.42, 2.50, 1.09, 0.37, 0.31, 0.10 and
0.04 mmol l–1, respectively. It thus appears that each
dextran has a characteristic cryoprotection profile. The
data show that the cryoprotecting efficacy and optimum
cryoprotecting concentrations of dextrans are inversely
related to their molecular mass. Dextran also served as a
significant cryoprotectant in the presence of glycerol (0.87
mol l–1) and dimethyl sulphoxide (0.76 mol l–1), which are
well known cryoprotectants; the action of the combined
cryoprotectants was almost additive. The presence of
glycerol or glycerol plus dimethyl sulphoxide caused a
significant reduction (from 8.42 to 6.27 mmol l–1) in the
optimum concentration of dextran. In the presence of the
three cryoprotectants, recovery of sperm motility was as
high as 58% (forward motility) and 60% (total motility).
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| Date |
2002
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| Type |
Article
PeerReviewed |
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| Format |
application/pdf
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| Identifier |
http://www.eprints.iicb.res.in/787/1/REPRODUCTION_123(_6)907%2D913;2002[53].pdf
Kundu, C N and Chakrabarty, J and Dutta, P and Bhattacharyya, D and Ghosh, A and Majumder, Gopal Chandra (2002) Effect of Dextrans on Cryopreservation of Goat Cauda Epididymal Spermatozoa using a Chemically Defined Medium. Reproduction, 123 (6). pp. 907-913. |
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| Relation |
http://dx.doi.org/10.1530/rep.0.1230907
http://www.eprints.iicb.res.in/787/ |
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