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Sains Malaysiana 48(9)(2019):
1937–1946
http://dx.doi.org/10.17576/jsm-2019-4809-15
Effect of
N-Acetylcysteine Supplementation on Oxidative Stress-Mediated Cryoinjury of
Bone Marrow Derived-Hematopoietic Stem Cells
(Kesan Suplementasi
N-Asetilsistein ke atas Kecederaan Krio Aruhan Tekanan Oksidatif pada Sel
Tunjang Hematopoetik Pencilan Sumsum Tulang)
SHAWAL MARADONA ABDUL WAHAB, ZARIYANTEY ABD HAMID*, RAMYA DEWI MATHIALAGAN
& IZATUS SHIMA TAIB
Biomedical
Science Programme, Centre for Health & Applied Sciences, Faculty of Health
Sciences, Universiti Kebangsaan Malaysia, Jalan Raja Abdul Muda Aziz, 50300
Kuala Lumpur, Federal Territory, Malaysia
Diserahkan: 18
Februari 2019/Diterima: 19 Jun 2019
ABSTRACT
Hematopoietic stem
cells (HSCs) transplantation was introduced as curative
treatment for various diseases. Cryopreservation of HSCs
is crucial for long term storage and maintenance of cellular quality; however,
it has been reported that cryopreservation itself causes oxidative
stress-driven apoptosis and cell loss. This study investigated impact of
supplementing N-acetylcysteine (NAC) as antioxidant during
cryopreservation on viability and oxidative stress in HSCs. HSCs
were isolated from murine bone marrow, cultured in HSCs
growth media and cryopreserved (1×106 cells per vial) together with
10% DMSO and NAC (0, 0.25, 0.5 or 2.0
μM) for 48 h, 2 weeks or 8 weeks at -196°C using controlled-rate-freezing
technique. Cell viability and oxidative stress in cryopreserved cells were
analysed at each time-point. Cell viability was significantly reduced (p<0.05) following cryopreservation as compared to
pre-cryopreservation. NAC supplementation significantly
increased cell viability (p<0.05) after 48 h cryopreservation at 0.5
μM and 2.0 μM and after 2 weeks cryopreservation at 0.25 μM
compared to the controls. Cryopreservation significantly enhanced GSH level
(p<0.05) and reduced MDA level (p<0.05)
without affecting SOD activity and PC level
in HSCs compared to pre-cryopreservation. NAC supplementation
significantly increased GSH level at 0.25 μM in
cryopreserved HSCs compared to control. In conclusion, NAC supplementation
during cryopreservation showed potential in minimizing cryoinjury by promoting
cell viability, increasing antioxidant capacity and reducing oxidative damage
in HSCs, however these effects are influenced by both
durations of cryopreservation and NAC concentration.
Keywords:
Cryopreservation; hematopoietic stem cells; N-acetylcysteine; oxidative stress
ABSTRAK
Pemindahan sel stem hematopoietik
(SSH) telah diperkenalkan sebagai rawatan
pelbagai penyakit. Pengawetan krio SSH adalah
penting untuk penyimpanan dan pemeliharaan kualiti SSH,
namun pengawetan krio dilaporkan berupaya mengaruh apoptosis dan
penurunan keviabelan sel akibat tekanan oksidatif. Kajian ini telah
mengkaji kesan suplementasi N-asetilsistien (NAC) sebagai antioksidan semasa
pengawetan krio terhadap keviabelan dan tekanan oksidatif SSH.
SSH diisolasi daripada sum-sum tulang mencit, dikulturkan di dalam
media pengkulturan SSH
dan seterusnya diawet krio (1×106 sel
per vial) dengan penambahan 10% DMSO dan NAC (0
μM, 0.25 μM, 0.5 μM atau 2.0 μM) selama 48 jam,
2 minggu atau 8 minggu pada suhu -196°C. Keviabelan sel dan
tekanan oksidatif ditentukan ke atas SSH pada setiap tempoh pengawetan krio. Keviabelan sel menurun
secara signifikan (p<0.05) selepas pengawetan
krio berbanding dengan pra-pengawetan krio. Suplementasi NAC
telah meningkatkan keviabelan sel secara ketara (p<0.05)
selepas pengawetan krio selama 48 jam pada kepekatan 0.5 μM
dan 2.0 μM dan juga selepas 2 minggu pengawetan krio pada kepekatan
0.25 μM berbanding kawalan. Pengawetan krio memberi kesan peningkatan
aras GSH (p<0.05)
dan penurunan aras MDA (p<0.05) tanpa memberi
kesan yang signifikan terhadap aras SOD dan PC berbanding
pra-pengawetan krio. Suplementasi NAC meningkatkan aras GSH
secara signifikan (p<0.05) pada kepekatan
0.25 μM dalam SSH yang
dikrio awet berbanding kawalan. Secara kesimpulannya, suplementasi
NAC
semasa pengawetan krio berpotensi meningkatkan keviabelan
sel dan kapasiti antioksidan serta mengurangkan kesan kerosakan
oksidatif, tetapi kesan ini adalah dipengaruhi oleh jangka masa
pengawetan krio dan kepekatan NAC yang
digunakan.
Kata kunci: N-asetilsistien; pengawetan krio; sel stem hematopoietik;
tekanan oksidatif
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*Pengarang untuk
surat-menyurat; email: zyantey@ukm.edu.my
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