title: Magnetic-activated cell sorting before cryopreservation preserves mitochondrial integrity in human spermatozoa.
abstract: Superparamagnetic annexin-V conjugated microbeads are able to eliminate spermatozoa with externalized phosphatidylserine, a membrane feature of apoptotic cells as well as spermatozoa with deteriorated plasma membrane. Our objective was to evaluate the effects of annexin-V Magnetic-Activated Cell Sorting (MACS) in cryopreservation-thawing protocols and on integrity of sperm mitochondrial transmembrane potential and mitochondrial integrity survival rate (MSR). Mature spermatozoa of 10 healthy donors were prepared by density gradient centrifugation and divided into 2 aliquots afterwards. The first one was subjected to annexin-V MACS followed by cryopreservation and thawing, while the second was cryopreserved-thawed without MACS to serve as control. Annexin-negative sperm separated by MACS showed significantly higher levels of intact mitochondria following cryopreservation-thawing (45.4+/-8.6%) compared to sperm that were not separated (15.8+/-4.6%, p<0.01). Separating a distinctive population of non-apoptotic spermatozoa with intact membranes may optimize cryopreservation-thawing outcome. MACS using annexin-V microbeads enhances the percentage of spermatozoa with intact transmembrane mitochondrial potential and mitochondrial integrity survival rates following cryopreservation.
authors.affiliation: Department of Dermatology/Andrology Unit, University of Leipzig, Germany.
authors.forename: Sonja
authors.initials: S
authors.lastname: Grunewald
authors.forename: Uwe
authors.initials: U
authors.lastname: Paasch
authors.forename: Tamer M
authors.initials: TM
authors.lastname: Said
authors.forename: Manja
authors.initials: M
authors.lastname: Rasch
authors.forename: Ashok
authors.initials: A
authors.lastname: Agarwal
authors.forename: Hans-Juergen
authors.initials: HJ
authors.lastname: Glander
mesh_terms: D017304:Annexin A5 / Q000378:metabolism; D002469:Cell Separation / Q000379:methods*; D015925:Cryopreservation / Q000379:methods*; D006801:Humans; D008280:Magnetics; D008297:Male; D008564:Membrane Potentials; D008928:Mitochondria / Q000502:physiology*; D013094:Spermatozoa / Q000166:cytology*; D014019:Tissue Donors
chemical_list: D017304:Annexin A5