title: Cryopreservation of very low numbers of spermatozoa from male patients undergoing infertility treatment using agarose capsules.
abstract: This study tried to cryopreserve low numbers of spermatozoa from men undergoing infertility treatments by inserting into agarose capsules. The capsules were transferred into a drop of cryoprotectant solution and injected 3-4 motile spermatozoa that were selected by the swim-up method by conventional intracytoplasmic sperm injection. These capsules were put on a Cryotop®and frozen in liquid nitrogen vapor, and then submerged into liquid nitrogen and subsequently thawed and recovered. The motile spermatozoa in the capsules were counted. Eventually, we cryopreserved 2142 motile spermatozoa in 702 agarose capsules from 26 male patients and 1356 (63%) spermatozoa maintained their motility after thawing. The spermatozoa motility rates after thawing (MRAT) ranged from 20.0% (5/25) to 95.1% (58/61) among patients. The median MRAT was 68.3% (interquartile range 46.1-75.7). The total number of motile spermatozoa collected by swim-up method strongly correlated with MRAT (r = 0.746). It was possible to cryopreserve spermatozoa from male patients undergoing infertility treatment using agarose capsules. However, there were wide differences in MRAT among patients. It seems the spermatozoa from semen where there were many motile spermatozoa may have higher freezing resistance. Further studies using this method in cryptozoospermic semen, testicular and epididymal spermatozoa are required.
authors.affiliation: Tokuoka Ladies Clinic, 1-3-1 Nakane, Meguro-Ku, Tokyo, 152-0031, Japan. hatakeyamas@infoseek.jp.|Graduate School of Health Science, Gumma Paz College, 1-7-1 Tonyamachi, Takasaki, 370-0006, Japan. hatakeyamas@infoseek.jp.|Yanaihara Women's Clinic, 1-26-29 Ofuna, Kamakura, 247-0056, Japan. hatakeyamas@infoseek.jp.
authors.forename: Shota
authors.identifier: http://orcid.org/0000-0002-8317-4713
authors.initials: S
authors.lastname: Hatakeyama
authors.affiliation: Tokuoka Ladies Clinic, 1-3-1 Nakane, Meguro-Ku, Tokyo, 152-0031, Japan.
authors.forename: Susumu
authors.initials: S
authors.lastname: Tokuoka
authors.affiliation: Graduate School of Science and Engineering, Yamagata University, 4-3-16 Jonan, Yonezawa, 992-8510, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Abe
authors.affiliation: The Institute for Advanced Reproductive Medical Technology, 909-21 Ishi, Fujimi-Cho, Maebashi, 371-0105, Japan.
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Araki
authors.affiliation: Graduate School of Health Science, Gumma Paz College, 1-7-1 Tonyamachi, Takasaki, 370-0006, Japan.|The Institute for Advanced Reproductive Medical Technology, 909-21 Ishi, Fujimi-Cho, Maebashi, 371-0105, Japan.|Nippon Reprogenetics, Maebashi, 909-21 Ishi, Fujimi-Cho, Maebashi, 371-0105, Japan.
authors.forename: Yasuhisa
authors.initials: Y
authors.lastname: Araki
mesh_terms: D002214:Capsules; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents; D006801:Humans; D007248:Infertility, Male / Q000628:therapy*; D008297:Male; D012685:Sepharose; D012996:Solutions; D020554:Sperm Injections, Intracytoplasmic / Q000379:methods; D013081:Sperm Motility; D013094:Spermatozoa / Q000502:physiology
chemical_list: D002214:Capsules; D003451:Cryoprotective Agents; D012996:Solutions; D012685:Sepharose
keywords: Agarose capsule; Cryopreservation; Human spermatozoa; ICSI; Motility rate