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Sains Malaysiana 52(11)(2023): 3103-3119
http://doi.org/10.17576/jsm-2023-5211-07
The Optimal Cryo Revival Period of
Cryopreserved Wharton's Jelly Derived-Mesenchymal Stem Cells
(Tempoh Pemulihan Krio Optimum bagi Sel Stem Mesenkima Wharton Jeli yang Diawet Krio)
VIKNESWARY RAVI KUMAR1,2, MOHAMAD FIKERI ISHAK2, SHARIFAH IZWAN TUAN
OTHMAN1, TRISTAN TAN3 & YOGESWARAN LOKANATHAN2,*
1University Selangor (UNISEL, Shah Alam City Campus, Selangor), Jalan Zirkon A7/A, Seksyen 7, 40000 Shah Alam, Selangor, Malaysia
2Centre for Tissue Engineering and
Regenerative Medicine, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Yaacob Latiff, Bandar Tun Razak, 56000 Cheras, Kuala Lumpur, Malaysia
3Da Vinci Healife Sdn. Bhd. 13-2, The Boulevard Offices, Mid Valley
City, 59200 Kuala Lumpur, Malaysia
Diserahkan: 12 Mei 2023/Diterima:
20 Oktober 2023
Abstract
The workflow of cryopreservation is a
challenging step in the standardised preparation of cell therapy products in
terms of methods used (i.e., adapted controls of the work environment, quality
control, reagents, and equipment). This study aimed to determine the effect of
cryopreservation on the stability of mesenchymal stem cell (MSC)
characteristics by comparing fresh cells with those that underwent different
post-thaw cell recovery periods. The MSCs were derived from human Wharton’s
jelly umbilical cord (n = 4). The cells that were cryopreserved for 7 days were
revived at 0 h (CRC-0h), 24 h (CRC-24h) and 7 days (CRC-7d) and then evaluated
on the basis of cell viability, doubling time, morphology, trilineage differentiation potential, growth kinetics, and MSC surface marker expression.
The cell viability of the CRC-0h group was 90%, whereas that of the CRC-24h group was 80%-85%. Cell attachment results showed that
CRC-24h had a notably higher attachment rate than the other two groups. The CRC
groups showed CD90, CD73 or CD44 expression, which meets the minimum criteria
for defining multipotent MSCs. By contrast, CD105 expression was significantly
reduced in the CRC groups and was lower than the minimum requirement based on
the standards of the International Society for Cellular Therapy. Results suggest
that at least 24 h is necessary to improve the quality of MSCs for it to be
adequate for cell therapy use.
Keywords: Cell therapy; cryopreservation; cryorevival; mesenchymal stromal cell; recovery period
Abstract
Aliran kerja pengawetan krio ialah satu langkah yang mencabar dalam penyediaan piawai produk terapi sel daripada segi kaedah yang digunakan (seperti kawalan disesuaikan persekitaran kerja, kawalan kualiti, reagen dan peralatan). Kajian ini bertujuan untuk menentukan kesan pengawetan krio ke atas kestabilan ciri sel stem mesenkima (MSC) dengan membandingkan sel segar dengan sel yang menjalani tempoh pemulihan sel pasca nyahbeku yang berbeza. MSC diperoleh daripada tali pusat jeli Wharton manusia (n = 4). Sel yang diawet krio selama 7 hari telah dihidupkan semula pada 0 jam (CRC-0j), 24 jam (CRC-24j) dan 7 hari (CRC-7h) dan kemudian dinilai berdasarkan daya maju sel, masa gandaan, morfologi, potensi pembezaan trilineage, kinetik pertumbuhan dan ekspresi penanda permukaan MSC. Daya maju sel kumpulan CRC-0j ialah 90%, manakala kumpulan CRC-24j ialah 80%–85%. Keputusan pelekatan sel menunjukkan bahawa CRC-24j mempunyai kadar pelekatan yang lebih tinggi daripada dua kumpulan lain. Kumpulan
CRC menunjukkan ekspresi CD90, CD73 atau CD44 yang memenuhi kriteria minimum untuk menentukan MSC multipoten. Sebaliknya, ekspresi CD105 berkurang dengan ketara dalam kumpulan CRC dan lebih rendah daripada keperluan minimum berdasarkan piawaian International Society for Cellular Therapy. Keputusan menunjukkan bahawa sekurang-kurangnya 24 jam diperlukan untuk meningkatkan kualiti MSC supaya ia mencukupi untuk kegunaan terapi sel.
Kata kunci: Pemulihan krio; pengawetan krio; sel stromal mesenkima; tempoh pemulihan; terapi sel
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*Pengarang untuk surat-menyurat; email:
lyoges@ppukm.ukm.edu.my
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