Safety of cryopreservation straws for human gametes or embryos: a preliminary study with human immunodeficiency virus-1.
The aim of this preliminary experimental study was to test the stability of cryopreservation straws to human immunodeficiency virus-1 (HIV-1). Three kinds of straws were tested: four polyvinyl chloride (PVC), four polyethylene terephthalate glycol (PETG) and 20 high-security ionomeric resin (IR). The PVC and PETG straws were sealed ultrasonically, and the IR straw by thermosoldering. Each sealed straw was cut in half to produce two demi-straws and then filled with 100 microl of HIV-1-containing supernatant (reverse transcriptase activity: 15 000 c.p.m./50 microl). The unsealed cotton end of PVC and PETG straws and the two halves of the IR straws (cotton and plastic plug ends) were tested. Each demi-straw was two- thirds submerged in RPMI medium at 37 degrees C, and RPMI samples were withdrawn on days 3, 7 and 11. Viral RNA was extracted from the medium and then amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) followed by nested PCR using primers specific to HIV-1 protease. On day 7, no HIV-1 RNA was detected in any of the different samples of medium that had surrounded the unsealed PVC and PETG straws with cotton ends, but three IR specimens were positive. On day 11, PVC and PETG remained negative but HIV-1 RNA was detected in RPMI samples for two more IR demi-straws (n = 5). In conclusion, under these experimental conditions (at 37 degrees C), the unsealed cotton end PVC, PETG and thermosoldered cotton end IR demi-straws appeared to be safe for HIV-1, while IR straws, sealed or unsealed with a plastic plug and with unsealed cotton ends, leaked.
Benifla
J L
JL
Department of Obstetrics and Gynaecology, Hôpital Bichat, 75018 Paris, France. jean-louis.benifla@bch.ap-hop-paris.fr
Letur-Konïrsch
H
H
Collin
G
G
Devaux
A
A
Kuttenn
F
F
Madelenat
P
P
Brun-Vezinet
F
F
Feldmann
G
G
D015925:Cryopreservation / Q000295:instrumentation* / Q000379:methods*; D004622:Embryo, Mammalian / Q000821:virology; D005854:Germ Cells; D015497:HIV-1; D006801:Humans; D011143:Polyvinyl Chloride; D012367:RNA, Viral / Q000032:analysis
D012367:RNA, Viral; D011143:Polyvinyl Chloride