Aplikasi Krioprotektan Ekstraseluler Tunggal Secara Efektif Mempertahankan Kualitas Sperma Manusia Pascavitrifikasi

Rini Widyastuti, Mas Rizky A. A. Syamsunarno, Mohammad Ghozali

Abstract


Pemilihan jenis krioprotektan merupakan salah satu kunci keberhasilan dalam mempertahankan motilitas dan viabilitas sperma pascavitrifikasi. Secara konvensional, vitrifikasi menggunakan konsentrasi krioprotektan dan laju kecepatan pembekuan yang tinggi untuk menghindari pembentukan kristal es intra dan ekstraseluler yang menyebabkan kerusakan dan kematian pada sel. Berdasar atas kemampuan menembus membran sel, krioprotektan dibedakan menjadi krioprotektan ekstra dan intraseluler. Sperma manusia memiliki struktur morfologi yang sangat padat dan sedikit mengandung sitoplasma sehingga perpindahan cairan selama proses vitrifkasi sangat kecil. Selain itu, sperma manusia juga mengandung beberapa jenis protein yang dapat berfungsi sebagai krioprotektan intraseluler. Berdasar atas kondisi tersebut, penggunaan krioprotektan pada vitrifkasi sperma manusia memerlukan studi lebih lanjut. Penelitian ini dilakukan di Laboratorium Sentral Universitas Padjadjaran dan dilaksanakan pada bulan Desember 2017–Januari 2018.Penelitian ini bertujuan mengetahui motilitas dan viabilitas sperma pascavitrifikasi dengan menggunakan tipe krioprotektan yang berbeda dan kombinasi keduanya. Earle’s balanced salt solution digunakan sebagai krioprotekan ekstraseluler, sementara EG 0,57% sebagai krioprotektan intraseluler. Sampel yang telah ditambahkan medium vitrifikasi diequilibrasi selama 10 menit, kemudian dikemas di dalam straw 0,25 mL dan langsung dipaparkan ke dalam nitrogen cair. Evaluasi dilakukan dengan melakukan thawing setelah 24 jam penyimpanan. Hasil menunjukkan bahwa motilitas dan viabilitas sperma tertinggi pada kelompok yang menggunakan media vitrifikasi krioprotektan ekstraseluler, walaupun secara statistik tidak berbeda nyata dengan kelompok lainnya (34%; 50%; p<0,05). Sebagai simpulan, krioprotektan ekstraseluler merupakan media vitrifikasi terbaik untuk menjaga motilitas dan viabilitas sperma pascavitrifikasi

Kata kunci: Krioprotektan, sperma manusia, vitrifikasi

 

Single Extracellular Cryoprotectant Application Effectively Maintain Post-Vitrification Human Sperm Quality

Selection of cryoprotectant is one of the keys to maintain sperm motility and viability after vitrification. Conventionally, vitrification uses cryoprotectants with high concentration and cooling rate to avoid the formation of intra- and extra-cellular ice crystals that can induce cell damage and cell death. Morphology structure of human sperm is very dense and contain less cytoplasm compartment; therefore mobilization of fluid is minimum during vitrification. In addition, human sperm also contains several types of protein that function as intracellular cryoprotectants. Based on this condition, the use of cryoprotectants in human sperm vitrification needs a further study. This study was conducted at the Central Laboratory of Universitas Padjadjaran in December 2017–January 2018. The aim was to determine the motility and viability of post-vitrification sperm by using different types of cryoprotectants. Samples that had been mixed with vitrification medium were equilibrated for 10 minutes, packed in a 0.25 mL straw, and directly exposed to liquid nitrogen. The evaluation was conducted  by thawing after 24 hours of storage. The results showed that the highest sperm motility and viability was  found in the group that used extracellular cryoprotectant vitrification media although it was not statistically different ( 34%; 50%, p <0.05) In conclusion, extracellular cryoprotectants are the best vitrification medium for maintaining motility and viability of post-vitrification sperm.

Key words: Cryoprotectant, sperm, vitrification

 


Keywords


Cryoprotectant; sperm; vitrification

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DOI: http://dx.doi.org/10.15395/mkb.v50n4.1319

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