Characteristics of Human Spermatozoa Harvested in Culture Media with and Without Serum Proteins

Abstract

This study was aimed to determine the efficiency of synthetic protein-free media in spermatozoa washing, preparation and retention of the activity of washed spermatozoa over short periods in vitro. Normozoospermic semen samples (n = 71) were equally apportioned and washed using synthetic protein-free medium (PFM), minimum essential medium + HSA (MEM) or commercial protein-containing medium (CPC). Washed spermatozoa were cultured in vitro using PFM, MEM or CPC media and held for 24 hrs at 4°C, 15°C, 22°C or 37°C. Spermatozoa activity was evaluated at 0 hr, 4 to 7 hrs and 24 hrs post-wash. The effects of PFM on spermatozoa motility, vitality, membrane integrity and DNA fragmentation level were not significantly different from that of MEM and CPC media at 0 hr, 4 to 7 hrs and 24 hrs post-wash in vitro. Synthetic PFM, MEM and CPC retained spermatozoa activity highest when specimen were held at 22°C and it was significantly higher (p < 0.05) than that at 37°C after 24 hrs incubation in vitro. However, no significant changes (p > 0.05) were noted in spermatozoa DNA fragmentation (SDF) levels when specimen were held at 22°C or 37°C at 4 to 7 hrs and also after 24 hrs post-wash in vitro in all media. The use of synthetic PFM as an alternative to the commercial protein-containing media in human spermatozoa washing and preparation procedure for an efficient and safer (Assisted Reproduction Technology) ART outcome. Spermatozoa activity can be successfully retained at room temperature post-wash over short periods; spermatozoa may lose viability rapidly if held for long hours at 37°C in all media. ABSTRAKKajian ini bertujuan untuk menentukan keberkesanan media bebas protein sintetik bagi pencucian spermatozoa, penyediaan dan penahanan aktiviti spermatozoa cucian dalam jangka masa terhad, secara in vitro. Sampel semen normozoospermik (n = 71) dibahagi-bahagikan secara sama rata dan dicuci dengan menggunakan medium bebas protein sintetik (PFM), medium esensial minimum + HSA (MEM) ataupun medium terkandung protein komersial (CPC). Spermatozoa cucian dikultur secara in vitro dengan menggunakan media PFM, MEM atau CPC dan ditahan selama 24 jam pada suhu 4°C, 15°C, 22°C atau 37°C. Aktiviti spermatozoa telah dinilai pada sela masa pascacucian 0 jam, 4 hingga 7 jam dan 24 jam. Kesan PFM terhadap motiliti, kevitalan, integriti membran dan tahap fragmentasi DNA tidak menunjukkan perbezaan signifikan berbanding kesan media MEM dan CPC pada sela masa 0 jam, 4 hingga 7 jam dan 24 jam pascacucian secara in vitro. PFM sintetik, MEM dan CPC mengekalkan aktiviti tertinggi bagi spermatozoa apabila spesimen ditahan pada suhu 22°C, dan aktiviti ini secara signifikan adalah lebih tinggi (p < 0.05) pada suhu 37°C selepas inkubasi secara in vitro selama 24 jam. Walau bagaimanapun, tiada perubahan signifikan (p > 0.05) diperhatikan terhadap tahap fragmentasi DNA spermatozoa (SDF) apabila spesimen ditahan pada suhu 22°C atau 37°C antara 4 hingga 7 jam dan juga selepas 24 jam proses pascacucian in vitro dilakukan dalam semua media. Penggunaan PFM si...