Sains Malaysiana 54(8)(2025): 1985-1994

http://doi.org/10.17576/jsm-2025-5408-09

 

Evaluating the Efficacy of Cryopreservation Media for the Preservation and Short-Term Storage of Human Dermal Fibroblast

(Menilai Keberkesanan Media Pengkrioawetan untuk Pemeliharaan dan Penyimpanan Jangka Pendek Fibroblas Dermal Manusia)

 

TITHTEEYA RATTANACHOT1, NUR RASYIDAH HAZIMAH MOHD ROSDI1, NUSAIBAH SALLEHUDDIN1, FAUZI MH BUSRA1,2 & MANIRA MAAROF1,2,3,*

 

1Department of Tissue Engineering and Regenerative Medicine, Faculty of Medicine, Universiti Kebangsaan Malaysia, 56000 Cheras, Kuala Lumpur, Malaysia

2Advance Bioactive Materials-Cells UKM Research Group, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor, Malaysia

3Ageing and Degenerative Disease UKM Research Group, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor, Malaysia

 

Diserahkan: 20 November 2024/Diterima: 17 Jun 2025

 

Abstract

Cryopreservation is a vital process for long-term preservation of cells without compromising their viability and functionality. Continuous cell culturing can lead to reduction in cell viability, higher risk of contamination, and increased reagent consumption. Optimizing short-term storage is essential to minimize cell damage and enhance cell adaptability for applications requiring brief storage duration. This study evaluates the effects of short-term storage on human dermal fibroblasts cryopreserved in different cryopreservation media. Redundant skin samples were obtained from surgeries with patient consent, processed, and sub-cultured to passage three (P3). Confluent cells were trypsinised and cryopreserved in three cryopreservation media: Foetal bovine serum with 10% dimethyl sulfoxide (FBS+10%DMSO), CryoStor10 (CS10), and cryo freezing serum-free media (CF-SFM). Cells were stored at -80 °C for 7 days, 14 days, and 1 month. Fibroblasts maintained their spindle-shaped, elongated morphology across all groups post-storage. The total number of live cells slightly decreased after 1 month, but no significant differences were found between the groups. Cell viability in CS10 after 1 month was significantly lower compared to the other storage durations, while no significant differences were observed in the other two media groups. Immunocytochemistry showed positive collagen type I (Col-1) and Ki67 expression at all storage durations. These findings suggest that fibroblasts retain their characteristics after short-term storage at -80 °C in different cryopreservation media. However, further studies are needed to examine the impact of long-term storage on other cell types.

Keywords: Cell characteristics; cryopreservation; fibroblasts; short-term storage

 

Abstrak

Pengkrioawetan merupakan proses penting untuk pemeliharaan jangka panjang sel tanpa menjejaskan daya hidup dan fungsinya. Kultur sel berterusan boleh menyebabkan pengurangan daya hidup sel, peningkatan risiko pencemaran dan penggunaan reagen yang tinggi. Pengoptimuman penyimpanan jangka pendek adalah penting untuk mengurangkan kerosakan dan meningkatkan kebolehsuaian sel bagi aplikasi yang memerlukan tempoh penyimpanan yang singkat. Penyelidikan ini menilai kesan penyimpanan jangka pendek terhadap fibroblas dermal manusia yang dikrioawet dalam media krioawetan berbeza. Sampel kulit berlebihan diperoleh daripada pembedahan dengan kebenaran pesakit, diproses dan dikultur sehingga subkultur 3 (P3). Sel yang mencapai konfluensi akan ditripsinkan dan dibekukan dalam tiga jenis media berbeza: serum janin lembu dengan 10% dimetil sulfoksida (FBS+10%DMSO), CryoStor10 (CS10) dan media bebas serum pembekuan kriogenik (CF-SFM). Sel ini disimpan pada suhu -80 °C selama 7 hari, 14 hari dan 1 bulan. Fibroblas mengekalkan morfologi berbentuk gelendong yang memanjang selepas penyimpanan. Jumlah keseluruhan sel hidup menurun sedikit selepas 1 bulan, namun tiada perbezaan yang ketara ditemui antara kumpulan. Keviabelan sel dalam CS10 selepas 1 bulan adalah lebih rendah berbanding tempoh penyimpanan yang lain, manakala tiada perbezaan diperhatikan dalam dua kumpulan media yang lain. Analisis imunositokimia menunjukkan ekspresi positif kolagen jenis I (Col-1) dan Ki67 pada semua tempoh penyimpanan. Penemuan ini menunjukkan bahawa fibroblas dapat mengekalkan cirinya selepas penyimpanan jangka pendek pada suhu -80 °C dalam media krioawetan berbeza. Namun, kajian lanjut diperlukan untuk menilai kesan penyimpanan jangka panjang pada jenis sel lain.

Kata kunci: Ciri sel; fibroblas; pengkrioawetan; penyimpanan jangka pendek

 

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*Pengarang untuk surat-menyurat; email: manira@ukm.edu.my

 

 

 

 

 

 

 

           

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