Investigation of Growth Properties of BHK-21 Cells with Different Characteristics to Comprise a Cell Bank in Low Serum-Supplement Medium in The Monolayer Culture System

Haydar Karadaş; Şükran Yılmaz; Muhammet Kaya

doi:10.18016/ksutarimdoga.vi.1759236

EN TR

Investigation of Growth Properties of BHK-21 Cells with Different Characteristics to Comprise a Cell Bank in Low Serum-Supplement Medium in The Monolayer Culture System

Abstract

BHK-21 cell lines with distinct growth characteristics are widely used in the production of Foot-and-Mouth Disease vaccines, and seed cell stocks are traditionally cultured in a monolayer system in GMEM medium containing 10% FBS. This study investigated the adaptation of BHK-21 An30 and BHK-21 An73 cell lines to Cellvento™ BHK-200 medium containing 2% FBS and examined the effects on their growth characteristics. Both cell lines underwent serial passages in serum-free medium supplemented with low serum, followed by analyses of viability, apoptosis, cell-cycle distribution, and colony-formation efficiency. Population doubling times were determined by generating growth curves after adaptation. BHK-21 An30 (t(33) = 2.410, p = .022*) and BHK-21 An73 (t(33) = 5.116, p < .001**) cultured in Cellvento™ BHK-200 medium showed higher growth than those grown in GMEM medium containing 10% FBS. Similarly, generation-dependent growth analyses demonstrated higher cell numbers for An30 (t(6) = 5.945, p = .012*) and An73 (t(6) = 2.882, p = .028*) in Cellvento™ BHK-200 compared with the control group. Cells grown in GMEM with 10% FBS exhibited higher colony-forming efficiency. Although viability and apoptosis rates did not differ significantly between the two media, the proportion of apoptotic cells decreased, and overall viability increased with additional passages in the low-serum-supplemented medium for both cell lines. The findings demonstrate that cell banks can be established in low-serum-supplemented, serum-free media, offering a feasible solution for large-scale cell culture production of Foot-and-Mouth Disease vaccines.

Keywords

Farklı Karakterdeki BHK 21 Hücrelerinin Düşük Serum Katkılı Vasattaki Büyüme Özelliklerinin Tek Tabaka Kültür Sisteminde İncelenmesi

Öz

Farklı büyüme karakteristiklerine sahip BHK-21 hücre hatları, Şap hastalığı aşılarının üretiminde yaygın olarak kullanılmaktadır. Tohum hücre stokları, monolayer kültür sisteminde %10 FBS içeren GMEM besiyerinde çoğaltılmaktadır. Bu çalışmada, BHK-21 An30 ve BHK-21 An73 hücre hatlarının %2 FBS içeren Cellvento™ BHK-200 besi ortamına adaptasyonu araştırılmış ve bu adaptasyonun hücre büyüme özellikleri üzerindeki etkisi incelenmiştir. Her iki hücre hattı da %2 FBS’li Cellvento™ BHK-200 besi ortamında seri pasajlara tabi tutulmuş ve ardından canlılık, apoptoz, hücre döngüsü ve koloni oluşturma verimliliği analiz edilmiştir. Popülasyonun iki katına çıkma süreleri belirlenmiş ve adaptasyon sonrası büyüme eğrileri oluşturulmuştur. %2 FBS’li Cellvento™ BHK-200 ortamında büyütülen BHK-21 An30 (t(33) = 2,410, p = .022*) ve BHK-21 An73 (t(33) = 5,116, p < .001**) hücre hatları, %10 FBS içeren GMEM ortamında büyütülenlere kıyasla daha yüksek büyüme göstermiştir. Benzer şekilde, jenerasyon bağımlı büyüme grafiklerinde Cellvento™ BHK-200 ortamında kültüre edilen An30 (t(6) = 5,945, p = .012*) ve An73 (t(6) = 2,882, p = .028*) hücrelerinin kontrol grubuna göre daha yüksek hücre sayılarına ulaştığı görülmüştür. %10 FBS içeren GMEM besiyerinde kültüre edilen hücreler ise daha yüksek koloni oluşturma verimliliği sergilemiştir. Hücre canlılığı ve apoptoz oranları iki besiyeri arasında anlamlı farklılık göstermemekle birlikte, düşük serum takviyeli ortamda ek pasajlarla her iki hücre hattında apoptotik hücre oranı azalmış ve canlılık artmıştır. Bulgular, düşük serum katkılı ortamlarda hücre bankalarının oluşturulabileceğini ve Şap hastalığı aşısı üretiminde büyük ölçekli hücre kültürleri elde etmek için uygulanabilir bir yaklaşım sunduğunu göstermektedir.

Anahtar Kelimeler

Thanks

The authors would like to thank the Directorate of the Foot and Mouth Disease Institute in Ankara for providing all the institute's facilities for the study. Haydar KARADAS performed this work in partial fulfillment of the M.Sc. degree in Agricultural Biotechnology at Eskişehir Osmangazi University, Turkiye. This study was presented at the 10th International Scientific Research Congress on Science and Engineering, held on April 11–12, 2021, in Ankara, Turkiye.

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Details

Primary Language

English

Subjects

Agricultural Biotechnology (Other)

Journal Section

Research Article

Authors

Haydar Karadaş
0000-0003-4481-448X
Türkiye

Şükran Yılmaz
0000-0002-7945-1124
Türkiye

Muhammet Kaya ^*
0000-0001-6474-121X
Türkiye

Early Pub Date

March 6, 2026

Publication Date

March 6, 2026

Submission Date

August 8, 2025

Acceptance Date

December 29, 2025

Published in Issue

Year 2026 Number: Advanced Online Publication

DOI

https://doi.org/10.18016/ksutarimdoga.vi.1759236

IZ

https://izlik.org/JA35XF96RL

Cite

RIS / Bibtex

APA

Karadaş, H., Yılmaz, Ş., & Kaya, M. (2026). Investigation of Growth Properties of BHK-21 Cells with Different Characteristics to Comprise a Cell Bank in Low Serum-Supplement Medium in The Monolayer Culture System. Kahramanmaraş Sütçü İmam Üniversitesi Tarım Ve Doğa Dergisi, Advanced Online Publication, 1147-1160. https://doi.org/10.18016/ksutarimdoga.vi.1759236