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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
Diserahkan: 22 Februari
2016/Diterima: 28 Jun 2016 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
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