Molecular Human Reproduction, Vol. 5, No. 1, 29-37,
January 1999
© 1999 European Society of Human Reproduction and Embryology
Nuclear chromatin variations in human spermatozoa undergoing swim-up and cryopreservation evaluated by the flow cytometric sperm chromatin structure assay
1 Section of Toxicology and Biomedical Sciences, ENEA CR Casaccia, Via Anguillarese 301, 00060 Rome, and 2 Laboratory of Seminology and Reproductive Immunology, Department of Medical Pathophysiology, University of Rome `La Sapienza', 00100 Rome, Italy
The sperm chromatin structure assay (SCSA) is a flow cytometric (FCM) technique which exploits the metachromatic properties of Acridine Orange to monitor the susceptibility of sperm chromatin DNA to in-situ acid denaturation. SCSA was used to study the chromatin structure variations of human spermatozoa in semen, both before and after swim-up and after cryopreservation. Semen samples were provided by 19 healthy normozoospermic subjects attending pre-marriage checks. Each sample was divided into three aliquots: the first aliquot was evaluated without further treatment, the second underwent swim-up, and the third was stored according to standard cryopreservation techniques in liquid nitrogen at –196°C. Samples were also analysed by light and fluorescence microscopy (after Acridine Orange staining to evaluate the number of green fluorescent sperm heads), and by computer-assisted semen analysis. The results showed that post-rise spermatozoa represent a subpopulation characterized by a general improvement of the morphological (reduction of the percentage of abnormal forms and heads, increase of the green head sperm percentage) and kinetic parameters. This subpopulation also exhibited improved chromatin structure properties, confirming that these cells have the best structural and functional characteristics, indicative of optimal fertilizing ability. On the other hand, overall sperm quality deteriorates after cryopreservation. When thawed spermatozoa underwent an additional swim-up round, a general improvement of nuclear maturity was seen in the post-rise spermatozoa.
cryopreservation/flow cytometry/human spermatozoa/sperm chromatin structure/swim-up
3 To whom correspondence should be addressed
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  M. Bungum, M. Spano, P. Humaidan, P. Eleuteri, M. Rescia, and A. Giwercman Sperm chromatin structure assay parameters measured after density gradient centrifugation are not predictive for the outcome of ART Hum. Reprod., January 1, 2008; 23(1): 4 - 10. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Gandini, F. Lombardo, D. Paoli, F. Caruso, P. Eleuteri, G. Leter, R. Ciriminna, F. Culasso, F. Dondero, A. Lenzi, et al. Full-term pregnancies achieved with ICSI despite high levels of sperm chromatin damage Hum. Reprod., June 1, 2004; 19(6): 1409 - 1417. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. I. Peris, A. Morrier, M. Dufour, and J. L. Bailey Cryopreservation of Ram Semen Facilitates Sperm DNA Damage: Relationship Between Sperm Andrological Parameters and the Sperm Chromatin Structure Assay J Androl, March 1, 2004; 25(2): 224 - 233. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Richthoff, M. Spano, Y.L. Giwercman, B. Frohm, K. Jepson, J. Malm, S. Elzanaty, M. Stridsberg, and A. Giwercman The impact of testicular and accessory sex gland function on sperm chromatin integrity as assessed by the sperm chromatin structure assay (SCSA) Hum. Reprod., December 1, 2002; 17(12): 3162 - 3169. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Lenzi and L. Gandini Characterization of human sperm Hum. Reprod., March 1, 2002; 17(3): 842 - 842. [Full Text] [PDF]
|
|
|
|
|
|
W. Kuczynski, M. Dhont, C. Grygoruk, D. Grochowski, S. Wolczynski, and M. Szamatowicz The outcome of intracytoplasmic injection of fresh and cryopreserved ejaculated spermatozoa--a prospective randomized study Hum. Reprod., October 1, 2001; 16(10): 2109 - 2113. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E.V. Younglai, D. Holt, P. Brown, A. Jurisicova, and R.F. Casper Sperm swim-up techniques and DNA fragmentation Hum. Reprod., September 1, 2001; 16(9): 1950 - 1953. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. T. Donnelly, E.K. Steele, N. McClure, and S. E.M. Lewis Assessment of DNA integrity and morphology of ejaculated spermatozoa from fertile and infertile men before and after cryopreservation Hum. Reprod., June 1, 2001; 16(6): 1191 - 1199. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Molina, J.A. Castilla, J.L. Castano, J. Fontes, N. Mendoza, and L. Martinez Chromatin status in human ejaculated spermatozoa from infertile patients and relationship to seminal parameters Hum. Reprod., March 1, 2001; 16(3): 534 - 539. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Sakkas, G.C. Manicardi, M. Tomlinson, M. Mandrioli, D. Bizzaro, P.G. Bianchi, and U. Bianchi The use of two density gradient centrifugation techniques and the swim-up method to separate spermatozoa with chromatin and nuclear DNA anomalies Hum. Reprod., May 1, 2000; 15(5): 1112 - 1116. [Abstract] [Full Text] [PDF]
|
|