title: Cryopreservation of human spermatozoa with minimal non-permeable cryoprotectant.
abstract: Cryopreservation of human spermatozoa is a commonly used technique in assisted reproduction, however freezing low concentrations of sperm while maintaining adequate post-thaw motility remains a challenge. In an effort to optimize post-thaw motility yields, low volumes of human sperm were frozen in polyimide-coated fused silica micro-capillaries using 0.065 M, 0.125 M, 0.25 M, or 0.5 M trehalose as the only cryoprotectant. Micro-capillaries were either initially incubated in liquid nitrogen vapor before plunging into liquid nitrogen, or directly plunged into liquid nitrogen. Post thaw sperm counts and motility were estimated. Spermatozoa that were initially incubated in liquid nitrogen vapor had greater post thaw motility than those plunged immediately into liquid nitrogen independent of trehalose concentration. The protective effect of 0.125 M d-glucose, 3-O-methyl-d-glucopyranose, trehalose, sucrose, raffinose, or stachyose were evaluated individually. Trehalose and sucrose were the most effective cryoprotectants, recovering 69.0% and 68.9% of initial sperm motility, respectively.
authors: Department of Surgery, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA; Shriners Hospital for Children, Boston, MA 02114, USA.
authors: Jie
authors: J
authors: Liu
authors: Department of Urology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA.
authors: Cigdem
authors: C
authors: Tanrikut
authors: Department of Obstetrics and Gynecology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA.
authors: Diane L
authors: DL
authors: Wright
authors: Department of Surgery, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA; Shriners Hospital for Children, Boston, MA 02114, USA.
authors: Gloria Y
authors: GY
authors: Lee
authors: Department of Surgery, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA; Shriners Hospital for Children, Boston, MA 02114, USA.
authors: Mehmet
authors: M
authors: Toner
authors: Department of Surgery, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA; Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA; Shriners Hospital for Children, Boston, MA 02114, USA. Electronic address: John_Biggers@hms.harvard.edu.
authors: John D
authors: JD
authors: Biggers
authors: Department of Obstetrics and Gynecology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA. Electronic address: TLTOTH@mgh.harvard.edu.
authors: Thomas L
authors: TL
authors: Toth
mesh_terms: D019325:3-O-Methylglucose / Q000494:pharmacology; D000818:Animals; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D005615:Freezing; D005947:Glucose / Q000494:pharmacology; D006801:Humans; D008297:Male; D009844:Oligosaccharides / Q000494:pharmacology; D011887:Raffinose / Q000494:pharmacology; D012662:Semen Preservation / Q000379:methods*; D013081:Sperm Motility / Q000187:drug effects* / Q000502:physiology*; D013094:Spermatozoa / Q000502:physiology*; D013395:Sucrose / Q000494:pharmacology; D014199:Trehalose / Q000494:pharmacology
chemical_list: D003451:Cryoprotective Agents; D009844:Oligosaccharides; D019325:3-O-Methylglucose; C005695:stachyose; D013395:Sucrose; D014199:Trehalose; D005947:Glucose; D011887:Raffinose
keywords: Cryoprotectant; Silica micro-capillary; Sperm cryopreservation; Sugars