title: On-chip cryopreservation: a novel method for ultra-rapid cryoprotectant-free cryopreservation of small amounts of human spermatozoa.
abstract: Cryopreservation of human spermatozoa free from cryoprotectant can avoid toxicity caused by highly concentrated cryoprotectant and a series of specific carriers have been previously explored, except for PDMS chip. Our study is aimed at exploring a novel device for ultra-rapid cryopreservation of small numbers of spermatozoa without cryoprotectant based on polydimethylsiloxane (PDMS) chips. Spermatozoa from 25 healthy men were involved in this study, comparing on-chip cryopreservation with different micro-channel height (group A: 10 µm height, group B: 50 µm height, group C: 100 µm height) and conventional freezing (group D) in liquid nitrogen for 72 h. The viability, motility, DNA integrity by comet assay and acrosome integrity by fluorescein isothiocyanate-conjugated peanut agglutinin (FITC-PNA) staining of frozen-thawed spermatozoa of each group were compared. The motility and viability of post-thawed spermatozoa was significantly decreased than that of pre-freezing spermatozoa. There was no difference of viability and motility of frozen-thawed spermatozoa between group A and D, while viability and motility of group B and C decreased compared to group A. Comet assay showed that no matter for group A or D, there was no difference of CR, TL, TD and OTM between pre-frozen and post-thawed spermatozoa. There was no difference of CR, TL, TD and OTM of post-thawed spermatozoa between group A and group D neither, while spermatozoa DNA damage was more serious in group B and group C with increasing height of micro-channel compared with group A. The proportion of intact acrosome of post-thawed spermatozoa in group A was the highest when compared with group B and group C, though similar to that of group D. In conclusion, PDMS chip with 10 µm height micro-channel is ideal for ultra-rapid cryopreservation of small quantity of spermatozoa without cryoprotectant.
authors: Reproductive Medicine Center, Renmin Hospital of Wuhan University, Wuhan, China.
authors: Yujie
authors: Y
authors: Zou
authors: Tailang
authors: T
authors: Yin
authors: Shijing
authors: S
authors: Chen
authors: Jing
authors: J
authors: Yang
authors: Weihua
authors: W
authors: Huang
mesh_terms: D000177:Acrosome / Q000502:physiology; D002470:Cell Survival; D020552:Comet Assay; D015925:Cryopreservation / Q000295:instrumentation / Q000379:methods*; D003451:Cryoprotective Agents; D005298:Fertility; D006801:Humans; D008297:Male; D015203:Reproducibility of Results; D012662:Semen Preservation / Q000295:instrumentation / Q000379:methods*; D013081:Sperm Motility; D013094:Spermatozoa / Q000166:cytology* / Q000502:physiology*
chemical_list: D003451:Cryoprotective Agents
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