Sains Malaysiana 46(3)(2017): 463–468

http://dx.doi.org/10.17576/jsm-2017-4603-14

Isolation Methods and Culture Conditions of Human Umbilical Vein Endothelial Cells from Malaysian Women

(Kaedah Pengasingan dan Keadaan Kultur Sel Endotelium Vena Umbilikus daripada Wanita Malaysia)

RAJA NOR SUHAILA & SABREENA SAFUAN*

School of Health Sciences, Universiti Sains Malaysia, 1610 Kubang Kerian, Kelantan Darul Naim

Malaysia

Received: 22 February 2016/Accepted: 28 June 20

ABSTRACT

Human umbilical vein endothelial cell (HUVEC) isolated from umbilical cord is widely used as endothelial cell model. However, HUVEC has been characteristically hard to maintain and showed molecular heterogeneity depending on the umbilical cord donors. Commercial HUVEC is commonly derived from European and Caucasian population which have different molecular characteristics from Asian women. This study aimed to optimize the isolation and culture condition of HUVEC using combinations of growth factors and extracellular matrix components so that the isolated HUVEC will purely represent the population under study. Umbilical cords were obtained from women post-labour. Different incubation times and digestive enzymes were used during endothelial cells isolation process. The culture conditions were optimized based on the coating materials and the media supplements. The results showed that 0.1% collagenase for 40 min incubation was the optimal isolation condition of HUVEC. HUVEC grown in 0.2% gelatin coated plate with 10% heat-inactivated fetal calf serum showed higher proliferative capacity and reduced cell death compared to other conditions (p<0.05). The results generated from this study provide a basic protocol of HUVEC isolation and culture conditions in order to generate working endothelial cell populations purely represent the Malaysian population.

Keywords: Endothelial cells; HUVEC; isolation; umbilical cord

ABSTRAK

Sel endotelium vena umbilikus manusia (HUVEC) yang dipencilkan daripada tali pusat digunakan secara meluas sebagai model sel endotelium. Walau bagaimanapun, HUVEC sukar untuk diselenggara dan menunjukkan keheterogenan molekul bergantung kepada penderma tali pusat. HUVEC komersial biasanya diperoleh daripada populasi Eropah dan Caucasian yang mempunyai ciri molekul yang berbeza daripada wanita Asia. Objektif kajian ini adalah untuk mengoptimumkan kaedah pengasingan dan keadaan kultur HUVEC menggunakan kombinasi faktor pertumbuhan dan komponen matriks ekstrasel supaya HUVEC yang dipencilkan ini akan mewakili populasi yang dikaji sahaja. Tali pusat diperoleh daripada wanita selepas bersalin. Masa pengeraman dan enzim pencernaan berbeza digunakan semasa proses pengasingan sel endotelium. Keadaan kultur dioptimumkan berdasarkan bahan salutan dan media tambahan. Keputusan menunjukkan bahawa 0.1% enzim kolagenase selama 40 minit tempoh pengeraman adalah keadaan pengasingan optimum HUVEC. HUVEC yang dikultur di atas 0.2% pinggan disalut gelatin dengan menggunakan 10% serum anak lembu yang diaktifkan haba menunjukkan kapasiti proliferatif yang lebih tinggi dan kurang kematian sel berbanding keadaan lain (p<0.05). Keputusan yang diperoleh daripada kajian ini menyediakan protokol asas mengenai pengasingan and keadaan kultur HUVEC untuk menjana populasi sel endotelium yang mewakili penduduk Malaysia secara khusus.

Kata kunci: HUVEC; pengasingan; sel endotelium; tali pusat

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*Corresponding author; email: sabreena@usm.my