Density Gradient Centrifugation Pra-freezing Mengoptimalkan Persentase Morfologi Normal Spermatozoa Pasca-thawing

Seso Sulijaya Suyono, Aucky Hinting, Hamdani Lunardhi, Reny I'tishom

Abstract


Kriopreservasi akan mengganggu struktur dan fungsi spermatozoa. Preparasi semen mampu menghasilkan spermatozoa dengan kualitas baik. Penelitian ini bertujuan menganalisis pengaruh preparasi semen dengan density gradient centrifugation (DGC) pra-freezing terhadap kualitas spermatozoa pasca-thawing. Penelitian dilakukan di boratorium Biologi Kedokteran, Fakultas Kedokteran Universitas Airlangga periode November 2017–Januari 2018. Penelitian eksperimental laboratoris dilakukan menggunakan cairan ejakulat volunter pria infertil. Semua sampel dibagi menjadi dua bagian, kelompok kontrol serta kelompok perlakuan berupa preparasi mini DGC. Setelah penambahan krioprotektan, selanjutnya dilakukan freezing. Pemeriksaan kualitas spermatozoa, meliputi motilitas, viabilitas serta persentase morfologi normal menggunakan metode WHO 2010, baik pra-freezing maupun pasca-thawing. Persentase perubahan kualitas spermatozoa pasca-thawing kedua kelompok dibandingkan menggunakan uji t dan bermakna jika nilai p<0,05. Total 20 sampel cairan ejakulat digunakan dalam penelitian ini. Persentase penurunan motilitas progresif, motilitas total, serta viabilitas pasca-thawing antara kedua kelompok tidak berbeda bermakna dengan nilai p masing-masing 0,422; 0,873 serta 0,432. Namun, penurunan persentase morfologi normal pasca-thawing pada kelompok kontrol jauh lebih besar daripada kelompok perlakuan dengan nilai p<0,001. Penelitian ini menyimpulkan bahwa preparasi semen berupa mini DGC pra-freezing mampu menghasilkan spermatozoa pasca-thawing dengan persentase morfologi normal yang lebih baik daripada protokol direct freezing.


Pre-freezing Density Gradient Centrifugation Optimizes the Percentage of Post-thawed Sperms with Normal Morphology

 Cryopreservation impairs sperm structure and functions. Sperm preparation is a selection technique to obtain a population of high quality sperms. This study aimed to analyze the effect of pre-freezing Density Gradient Centrifugation (DGC) sperm preparation on the quality of post-thawed sperms. An experimental laboratory study was conducted using the ejaculates collected from volunteers visiting to our department. Samples were split into two fractions: control group and treatment group. In the treated group, mini-DGC sperm preparations were developed. After some cryoprotectants were added, samples were then cryopreserved using rapid freezing protocol. The evaluation of sperm quality that included evaluation on motility, viability and percentage of sperm morphology was performed by referring to 2010 WHO standardization on semen analysis. Evaluation was performed under pre-freezing and post-thawed condition. The percentage of sperm parameter changes between the two groups were compared using t-test with p-value <0.05 considered statistically significant. A total of 20 samples were included in this study. Post-thawed progressive motility, total motility, and viability considerably declined between two groups with p-values of 0.422, 0.873, and 0.432 respectively. In post-thawed observation, the percentage of sperms with normal morphology was significantly lower in the control group when compared to the treatment group  (p<0.001). In conclusion, the pre-freezing mini-DGC can optimize the post-thawed percentage of sperms with normal morphology compared to direct freezing protocol


Keywords


Density gradient; freezing; spermatozoa; thawing

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DOI: http://dx.doi.org/10.15395/mkb.v50n3.1348

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