Journal of Uludağ University Medical Faculty

1300-414X 2645-9027

Bursa Uludağ University

10.32708/uutfd.1897295

Reproductive Medicine (Other)

Üreme Tıbbı (Diğer)

Kumulus Hücresi Ekstraselüler Matriksi ile Desteklenen Sperm Hazırlama Protokollerinin İn Vitro Sperm Kalitesi Üzerindeki Etkisi

Effects of Cumulus Cell Extracellular Matrix–Assisted Sperm Preparation Protocols on In Vitro Sperm Quality

https://orcid.org/0000-0003-2664-3066

Aghayeva

Sabina

Department of Histology and Embryology, Faculty of Medicine, Bursa Uludağ University, Bursa, Türkiye.

https://orcid.org/0009-0000-8217-4642

Durdu

Talha Yasin

Department of Histology and Embryology, Faculty of Medicine, Bursa Uludağ University, Bursa, Türkiye.

https://orcid.org/0000-0002-8332-7353

Çakır

Cihan

Department of Histology and Embryology, Faculty of Medicine, Bursa Uludağ University, Bursa, Türkiye.

https://orcid.org/0000-0002-9277-7735

Aslan

Kiper

Department of Obstetrics and Gynecology, Faculty of Medicine, Bursa Uludağ University, Bursa, Türkiye.

https://orcid.org/0000-0002-1953-2475

Kasapoğlu

Işıl

Department of Obstetrics and Gynecology, Faculty of Medicine, Bursa Uludağ University, Bursa, Türkiye.

https://orcid.org/0000-0003-4969-3584

Gök Yurtseven

Duygu

Department of Histology and Embryology, Faculty of Medicine, Bursa Uludağ University, Bursa, Türkiye.

03 26 2026

52 1897295 02 25 2026 03 24 2026

1975

Journal of Uludağ University Medical Faculty

Bu çalışmada, üremeye yardımcı tedavilerde (ÜYTE) uygulanan sperm hazırlama tekniklerinde kültür medyumuna kumulus hücresi ekstraselüler matriksi (CCECM) eklenmesinin; rutin sperm parametreleri, fertilizasyon kapasitesi ve sperm DNA fragmantasyonu üzerindeki etkilerini belirlemek ve sperm ayrıştırma protokollerini karşılaştırmalı olarak analiz etmek amaçlanmıştır. Çalışmada, normozoospermik 30 farklı erkek hastadan alınan ejakülat örnekleri ile bu hastaların eşlerinden toplanan kumulus-oosit kompleksleri kullanıldı. Her hastadan elde edilen semen örneği beş eşit hacme ayrılarak kontrol, dansite gradiyent (DG), dansite gradiyent + kumulus hücresi ekstraselüler matriksi (DG+K), swim-up (SU), swim-up + kumulus hücresi ekstraselüler matriksi (SU+K) olmak üzere beş grup oluşturuldu. Elde edilen sonuçlar, tüm deney gruplarında sperm konsantrasyonunun kontrol grubuna kıyasla belirgin olarak azaldığını; bu azalmanın özellikle swim-up uygulanan grupta anlamlı düzeye ulaştığını gösterdi. Motilite oranı en yüksek SU+K grubunda saptanırken, DG+K ve SU gruplarında da motilitenin anlamlı biçimde arttığı belirlendi. TPMSS değeri ise SU ve SU+K gruplarında anlamlı olarak azaldı. Morfoloji açısından gruplar arasında belirgin bir farklılık izlenmezken, canlılık oranının CCECM ilavesi yapılan gruplarda kontrol grubuna göre dikkat çekici şekilde arttığı görüldü. Akrozomal reaksiyon kapasitesini yansıtan ARIC skorları DG+K ve SU+K gruplarında anlamlı derecede yüksek bulundu. Ayrıca, DNA hasarını gösteren DFI değerleri bu iki grupta anlamlı olarak daha düşük saptandı ve CCECM katkısının sperm DNA bütünlüğünü koruyabileceği ortaya kondu. Sperm yıkama sırasında kullanılan kültür medyumuna CCECM eklenmesi, özellikle swim-up yöntemiyle birlikte uygulandığında sperm kalitesi ve verimliliğini artırdı. Bu yaklaşımın, ÜYTE kapsamında erkek infertilitesi tedavisinde uygulanabilir bir seçenek olabileceğini düşünüyoruz.

This study aimed to determine the effects of adding cumulus cell extracellular matrix (CCECM) to the culture medium used in sperm preparation techniques for assisted reproductive treatments (ART) on routine sperm parameters, fertilization capacity, and sperm DNA fragmentation, and to compare sperm separation protocols. Ejaculate samples from 30 normozoospermic male patients and cumulus–oocyte complexes collected from their spouses were included. Each semen sample was divided into five equal volumes and allocated to five groups: control, density gradient (DG), density gradient plus cumulus cell extracellular matrix (DG+K), swim-up (SU), and swim-up plus cumulus cell extracellular matrix (SU+K). The results showed that sperm concentration was markedly lower in all experimental groups than in the control group, with the decrease reaching statistical significance, particularly in the swim-up group. Motility was highest in the SU+K group and was also significantly increased in the DG+K and SU groups. In contrast, the TPMSS value decreased significantly in the SU and SU+K groups. No marked differences were observed among the groups in terms of morphology. Viability increased notably in the CCECM-supplemented groups, and this increase remained significant across all experimental groups compared with the control group. ARIC scores, reflecting acrosomal reaction capacity, were significantly higher in the DG+K and SU+K groups. Moreover, DFI values, indicating DNA damage, were significantly lower in these two groups, suggesting that CCECM supplementation may help preserve sperm DNA integrity. Adding CCECM to the culture medium used in sperm washing improved sperm quality and efficiency, particularly with the swim-up method. We believe this approach may be a feasible option for treating male infertility within assisted reproductive technologies (ART).

Cumulus cell extracellular matrix Sperm quality Density gradient Swim-up DNA fragmentation Acrosome reaction

Kumulus hücresi ekstraselüler matriksi Sperm kalitesi Dansite-gradiyent Swim-up DNA fragmantasyonu Akrozom reaksiyonu

Bursa Uludağ University Scientific Research Projects Coordination Unit (BAP)

TYL-2025-2129

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