Aktive Trombositten Zengin Plazmanın Testis Organoidlerinde In Vitro Spermatogeneze Etkisi

Abstract

İnfertilite, çiftlerin %10–20’sini etkiler ve erkek kaynaklı faktörler infertilite vakalarının %30–50’sinden sorumludur. Testis organoid (TO) kültür sistemleri in vitro spermatogenezi desteklese de, bu yaklaşımların en önemli limitasyonlarından biri kültür ortamlarının içeriği tam olarak bilinmeyen serum ve/veya farklı tür kökenli büyüme faktörleri ile desteklenmesidir. Trombositten zengin plazma (PRP), testiküler hasar sonrası üzerinde yenileyici etkisi sahiptir ve spermatogonyal kök hücrelerin (SKH) in vitro proliferasyonunu ve fonksiyonunu korur. Bu çalışmada, prepubertal C57BL/6 testiküler hücreler ultra düşük bağlanma ve hava–sıvı ara yüzey kültür sistemleri kullanılarak aktive PRP ile kültüre edilmiştir. Spermatogenez 1., 3., 4. ve 6. haftalarda histomorfometrik ve immünohistokimyasal analizlerle değerlendirilmiştir. %10 PRP ile desteklenen TO’larda (PRP grubu), SKH havuzu 6 hafta boyunca korunmuş ve ID4(+) SSC sayısı, 3., 4. ve 6. haftalarda %10 knockout serum replacement (KK grubu) ve %5 knockout serum replacement + %5 fetal sığır serumu (KF grubu) ile desteklenen TO’lara oranla daha yüksek bulunmuştur. PRP grubunda SCP3(+) spermatosit sayısı 1. haftadan 6. haftaya kadar sabit kalmış ve 4. ve 6. haftalarda KK ve KF gruplarına göre daha yüksek bulunmuştur. Tüm TO’larda 3. haftada ACR(+) yuvarlak spermatidler gözlemlenmiş ve 4. haftada PRP grubunda ACR(+) spermatid KK grubundan daha fazladır. Sonuçlar, aktifleştirilmiş PRP’nin spermatogenezi desteklemek için potansiyel bir kültür takviyesi olabileceğini, ancak güvenliği ve klinik uygulanabilirliğinin daha fazla araştırmaya ihtiyaç duyduğunu göstermektedir.

Keywords

• Erkek infertilitesi
• Testis organoid kültürü
• Aktive trombositten zengin plazma (PRP)
• İn vitro spermatogenez

Effect of Activated Platelet-Rich Plasma on In Vitro Spermatogenesis in Testicular Organoids

Abstract

Infertility affects 10–20% of couples, and male-related factors contribute to 30–50% of cases. Although testicular organoids (TOs) support in vitro spermatogenesis, one of the most significant limitations of these approaches is the undefined nature of serum and/or xenogenic growth factors used in the culture media. Platelet-rich plasma (PRP) has a regenerative effect on spermatogenesis after testicular damage and maintains the proliferation and function of spermatogonial stem cells (SSCs) in vitro. In this study, prepubertal C57BL/6 testicular cells were cultured with activated PRP using ultra-low attachment and air–liquid interface culture systems. Spermatogenic progression was evaluated at weeks 1, 3, 4, and 6 through histomorphometric and immunohistochemical analyses. In TOs supported with 10% PRP (PRP group), the SSC pool was preserved for 6 weeks, and the number of ID4(+) SSCs was higher than that in TOs supported with 10% knockout serum replacement (KK group) and 5% knockout serum replacement + 5% fetal bovine serum (KF group) at weeks 3, 4, and 6. In the PRP group, the number of SCP3(+) spermatocytes remained constant from weeks 1 to 6 and was higher than that in the KK and KF groups at weeks 4 and 6. In all TOs, ACR(+) round spermatids were observed at week 3, and PRP group had more ACR(+) spermatids than KK group at week 4. The results suggest that activated PRP may serve as a promising culture supplement to support spermatogenesis, although its safety and clinical applicability require further investigation.

Keywords

• Male infertility
• Testicular organoid culture
• Activated platelet-rich plasma (PRP)
• In vitro spermatogenesis

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