title: Vitrification of human ICSI/IVF spermatozoa without cryoprotectants: new capillary technology.
abstract: The aim of this study was to develop and to test the standardized aseptic  technology of permeable cryoprotectant-free vitrification of human spermatozoa  in capillaries (for intracytoplasmic sperm injection [ICSI] or in vitro  fertilization [IVF]). To test the effect of vitrification on basic sperm  parameters, each of 68 swim-up-prepared ejaculates from  oligo-astheno-terato-zoospermic patients were aliquoted and distributed into 3  groups: 1) nontreated control, 2) 10 μL of spermatozoa cryopreserved by  slow conventional freezing with glycerol-contented medium, and 3) 10 μL of  spermatozoa vitrified in 50-μL plastic capillaries in culture medium with  0.25 M sucrose. Spermatozoa motility (1, 24, and 48 hours after warming),  plasma membrane integrity, acrosomal integrity, and spontaneous  capacitation-like changes were determined after warming. Aseptic  cryoprotectant-free vitrification showed a significantly stronger  cryoprotective effect compared with conventional freezing. One hour after  warming, motility, plasma membrane integrity, and acrosomal integrity were  significantly higher than is observed for conventionally frozen spermatozoa  (28% vs 18%, 56% vs 22%, and 55% vs 21%, respectively; P < .05),  although lower than in fresh spermatozoa (35%, 96%, and 84%, respectively;  P < .05). Capacitation-like changes did not differ significantly  between vitrified and conventionally frozen samples (8% vs 9%, respectively;  P > .1) (2% in fresh spermatozoa). The newly developed technology  of aseptic vitrification of human spermatozoa in capillaries can effectively  preserve these cells from cryo-injures. Spermatozoa, vitrified by this  technology, are free from seminal plasma owing to swim-up preceding  vitrification and are free from permeable cryoprotectants. They are ready for  further use immediately after warming without any additional treatment.  Therefore, the reported technology has a great potential for use in  ICSI/IVF.
authors: Department of Obstetrics and Gynecology, Cologne University, Cologne, Germany. v.isachenko@yahoo.com
authors: V
authors: V
authors: Isachenko
authors: R
authors: R
authors: Maettner
authors: A M
authors: AM
authors: Petrunkina
authors: K
authors: K
authors: Sterzik
authors: P
authors: P
authors: Mallmann
authors: G
authors: G
authors: Rahimi
authors: R
authors: R
authors: Sanchez
authors: J
authors: J
authors: Risopatron
authors: I
authors: I
authors: Damjanoski
authors: E
authors: E
authors: Isachenko
mesh_terms: D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology; D006801:Humans; D008297:Male; D055101:Semen Analysis; D013075:Sperm Capacitation; D020554:Sperm Injections, Intracytoplasmic; D013094:Spermatozoa; D058989:Vitrification
chemical_list: D003451:Cryoprotective Agents