Sains Malaysiana 48(5)(2019): 1105–1110

http://dx.doi.org/10.17576/jsm-2019-4805-19

 

Optimization of Cryopreserved Single Cell Suspension of Gastric Epithelial Cells for Successful DotScanTM Antibody Microarray

(Pengoptimuman Pengkrioawetan Ampaian Sel Tunggal Sel Epitelium Gaster bagi Kejayaan Mikroatur Antibodi DotScanTM)

ASIF SUKRI1, ALFIZAH HANAFIAH1*, NIK RITZA KOSAI2, MUSTAFA MOHAMED TAHER2,

ISA MOHAMED ROSE3 & RAMELAH MOHAMED4

 

1Department of Medical Microbiology and Immunology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Yaacob Latiff, Bandar Tun Razak, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

 

2Department of Surgery, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Yaacob Latiff, Bandar Tun Razak, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

 

3Department of Pathology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Yaacob Latiff, Bandar Tun Razak, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

 

4Biotech Diagnostics, UKMTech, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor Darul Ehsan, Malaysia

 

Received: 9 September 2016/Accepted: 5 March 2019

 

ABSTRACT

Cell cryopreservation plays a pivotal role for successful immunological assays. DotScanTM antibody microarray employs live cells with satisfactory cell viability for detection of clusters of differentiation (CD) antigens in various diseases. Effective cryopreservation contributes to successful capture of cells on DotScanTM antibody microarray. The purpose of this study was to evaluate the effect of cells viability on cells binding to specific CD antigens. Cells with similar concentration but different percentage of cell viabilities were captured on DotScanTM slides. Cells stored for 5 days at -80ºC and added with freezing medium in stepwise manner had higher viabilities as compared to cells stored for 6 months at -80ºC and added with freezing medium in direct manner (p=0.012). Cells with 88% viability had higher binding to CD44/CD29 control antibodies as compared to cells with 33% viability. Short-term storage of cells and the technique for freezing medium addition to cells play a critical role for successful cryopreservation of single cell suspension.

 

Keywords: Cell surface antigens; cryopreservation; DotScanTM antibody microarray; epithelial cells

 

ABSTRAK

Pengkrioawetan sel memainkan peranan penting dalam kejayaan asai imunologi. Mikroatur antibodi DotScanTM memerlukan sel hidup yang memuaskan untuk pengesanan antigen kluster pembezaan (CD) dalam pelbagai penyakit. Keberkesanan pengkrioawetan menyumbang kepada kejayaan penangkapan sel ke atas mikroatur antibodi DotScanTM. Tujuan kajian ini adalah untuk menilai kesan daya hidup sel ke atas pelekatan sel terhadap antigen CD yang khusus. Sel dengan kepekatan yang sama tetapi daya hidup yang berlainan ditangkap ke atas slaid DotScanTM. Sel disimpan selama 5 hari pada suhu -80°C dan ditambah media takat beku secara berperingkat mempunyai daya hidup yang lebih tinggi berbanding sel yang disimpan selama 6 bulan dan ditambah media takat beku secara langsung (p=0.012). Sel dengan daya hidup 88% mempunyai pelekatan yang lebih tinggi terhadap antibodi kawalan CD44/CD29 berbanding sel dengan daya hidup 33%. Sel yang disimpan untuk jangka masa singkat dan teknik menambah media takat beku terhadap sel memainkan peranan penting untuk kejayaan pengkrioawetan ampaian sel tunggal.

 

Kata kunci: Antigen permukaan sel; mikroatur antibodi DotScanTM; pengkrioawetan; sel epitelium

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*Corresponding author; email: alfizah@ppukm.ukm.edu.my

 

 

 

 

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