Z.A. Intan Zarina

National University of Malaysia, Putrajaya, Putrajaya, Malaysia

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Publications (7)0 Total impact

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    ABSTRACT: Current haematopoietic stem cell transplantation protocols rely heavily upon CD34+ cells to estimate haematopoietic stem and progenitor cell yield. However, several studies nowadays emerged with evidence that CD133+ cells represent a more primitive cell population than their CD34+ counterparts. The objective of the present study was to isolate CD133+ stem cells by a rapid negative isolation method which depletes human peripheral blood mononuclear cells (MNCs) from cells expressing haematopoietic lineage markers CD2, CD3, CD11b, CD14,CD15, CD16, CD19, CD56, CD123, and CD235a and isolates a discrete lineage negative (Lin-) cell population (11±2% of MNC, n=6). Human peripheral blood MNCs were fractionated by density gradient centrifugation. After labelling with antibody against haematopoietic lineage markers, MNCs were subjected to magnetic assisted cell sorting (MACS). The expression of stem cells markers was examined by reverse transcriptase PCR (RT-PCR). We showed that Lin- stem-progenitor cell population encompassed the common markers used to characterize adult stem cells including SLAMF1, CD90 and CD133 and was also enriched for CD117, a surface receptor for stem cell factor. This cell population is more homogenous and primitive as compared to suspension MNCs which comprised of SLAMF1 and CD117 but the cells with haematopoietic lineage marker, CD14 and CD2 were also present. In conclusion, our approach was able to isolate a homogenous Lin- stem-progenitor cell population from human peripheral blood hence offers a promising alternative to current haematopoietic stem and progenitor selection methods.
    No preview · Article · Jun 2014 · Malaysian Applied Biology
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    ABSTRACT: Tujuan kajian ini adalah untuk mengenalpasti perhubungan diantara aktiviti spesifik laktat dehidrogenase (LDH), aspartat aminotransferase (AST), asid fosfatase rintang tartrat (TRAP) dan alkali fosfatase (ALP) dengan jumlah enzim masing-masing yang hadir dalam cecair krevis gingiva (gingiva crevicular fluid; GCF) semasa rawatan ortodontik dengan menggunakan analisis statistik. Sampel GCF diambil daripada 19 orang subjek ortodontik pada masa-masa tertentu; sebelum memakai pendakap gigi (basal) dan selepas memakai pendakap gigi [minggu 0 (sebelum dikenakan tekanan) dan minggu 1 hingga minggu 5 (selepas daya diberikan)]. Subjek menerima daya 1.0 N dan 1.5 N sama ada di bahagian kanan atau kiri arkus maksila. Pengasaian enzim dan analisis ELISA dilakukan. Analisis korelasi Pearson menunjukkan tiada korelasi yang tinggi dan signifikan (p>0.05) di antara jumlah enzim dengan aktiviti spesifik enzim semasa rawatan ortodontik pada kedua-dua daya (1.0 N dan 1.5 N) bagi kesemua enzim penanda yang dikaji; LDH (1.0 N, r=-0.28, p>0.05; 1.5 N, r=-0.08, p>0.05), AST (1.0 N, r=-0.49, p>0.05; 1.5 N, r=0.08, p>0.05), TRAP (1.0 N, r=0.016, p>0.05; 1.5 N, r=0.48, p>0.05) dan ALP (1.0 N, r=-0.28, p>0.05; 1.5 N, r=-0.08, p>0.05). Oleh itu, asai enzim lebih sesuai digunakan berbanding analisis ELISA untuk melihat profil aktiviti enzim penanda yang aktif semasa rawatan ortodontik berdasarkan kepada proses pergerakan gigi yang melibatkan enzim aktif.
    No preview · Article · Jun 2014 · Malaysian Applied Biology

  • No preview · Article · May 2014 · Journal of Applied Sciences
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    ABSTRACT: Stem cells that made up of embryonic stem cells and adult stem cells are well known for their ability to self renew and differentiate into matured cells. Adult stem cells isolated from peripheral blood system were used as the study sample. The objective of this study was to determine the relationship between size and proliferation potential of stem cells from 3 different culturing terms, i.e., short (7 days), medium (14 days), and long (30 days) terms. Density gradient centrifugation was applied in isolating mononucleated cells (MNCs) from peripheral blood sample. Isolated MNCs were washed with Hank's Balanced Salt Solution and Phosphate Buffer Saline. The MNCs which cultured in complete media were then subjected to proliferation analysis every day for a time period of 30 days. RT-PCR analyses and cell size measurements were carried out at day 7, 14, and 30. Activation of KIT and SLAMF1 marker genes during RT-PCR analyses indicated that the isolated MNCs were stem cells. Analyses of proliferation rate and cell size showed that short term stem cells have the largest cell size with the lowest proliferation potential. While, medium term stem cells gave rise to a smaller stem cell population with higher proliferation potential compared to short term stem cells. Long term stem cells have smallest cell size with highest proliferation potential. In conclusion, each culturing term cell population had their own sizes and can be isolated based on those sizes.
    No preview · Article · Dec 2013 · Malaysian Applied Biology
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    ABSTRACT: Haematopoietic stem cells (HSCs) are used in the therapy of blood disorders due to the ability of these cells to reconstitute haematopoietic lineage cells when transplanted into myeloablative recipients. However, substantial number of cells is required in order for the reconstitution to take place. Since HSCs present in low frequency, larger number of donor is required to accommodate the demand of transplantable HSCs. Therefore, in vitro expansion of HSCs will have profound impact on clinical purposes. The aim of this study was to expand lineage negative (Lin(-)) stem cells from human peripheral blood. Total peripheral blood mononuclear cells (PBMNCs) were fractionated from human blood by density gradient centrifugation. Subsequently, PBMNCs were subjected to magnetic assisted cell sorter (MACS) which depletes lineage positive (Lin(+)) mononuclear cells expressing lineage positive markers such as CD2, CD3, CD11b, CD14, CD15, CD16, CD19, CD56, CD123, and CD235a to obtained Lin(-) cell population. The ability of Lin(+) and Lin(-) to survive in vitro was explored by culturing both cell populations in complete medium consisting of Alpha-Minimal Essential Medium (AMEM) + 10% (v/v) Newborn Calf Serum (NBCS)+ 2% (v/v) pen/strep. In another experiment, Lin+ and Lin-were cultured with complete medium supplemented with 10ng/mL of the following growth factors: stem cell factor (SCF), interleukin (IL)-3, granulocyte-macrophage colony stimulating factor (GM-CSF), 2IU/mL of Erythropoietin (Epo) and 20ng/mL of IL-6. Three samples were monitored in static culture for 22 days. The expansion potential was assessed by the number of total viable cells, counted by trypan blue exclusion assay. It was found that Lin+ mononuclear cells were not able to survive either in normal proliferation medium or proliferation medium supplemented with cytokines. Similarly, Lin-stem cells were not able to survive in proliferation medium however, addition of cytokines into the proliferation medium support Lin-stem cells for at least 18 days. The Lin-stem cells started to response to the cytokines added as early as Day 2 of culture. It is concluded that Lin-stem cells can be expanded in vitro by culturing in proliferation medium supplemented with cytokines.
    Preview · Conference Paper · Nov 2013
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    ABSTRACT: Problem statement: DNA samples from fourteen modern human bloods (seven males and seven females) and two ancient skeletal samples excavated from Kalumpang Island and Dungun, Peninsular Malaysia were subjected for molecular genders determination using specific primers of human AMELX and AMELY . Approach: A standard multiple PCR mixture with forward primer and either a human X-specific reverse primer for AMELX or a human Y-specific reverse primer for AMELY amplifications were used to assess the presence of these genes in male and female samples. Results: PCR amplification of a modern male sample yielded 329 and 235 bp bands, whereas a modern female sample only 329 bp band. The Kalumpang Island sample produced two positive bands of AMELY and AMELX . After reamplification of the Dungun sample, only an AMELY band was visible. All amplified bands were cloned into TA plasmid and sequenced. BLAST analysis showed that the 329 bp band is AMELX , while the 235 bp product is AMELY . Conclusion/Recommendations: The skeletal remains of both Kalumpang Island and Dungun samples from west and east of Peninsular Malaysia respectively, are males.
    Full-text · Article · Oct 2009 · American Journal of Applied Sciences
  • Z.A. Intan Zarina · Z.A. Shahrul Hisham · Rohaya M.A.W · Sahidan S · Zaidah Z.A
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    ABSTRACT: The aim of this study is to differentiate peripheral blood mononucleated cells that have been isolated from ICR mice`s ( Mus musculus )peripheral blood into mature osteoblast and osteoclast cells with addition of growth factors. The mononucleated cells were isolated by density centrifugation using Ficoll-Paque<sup>TM</sup> Plus. The culture mediums were then supplemented with growth factors; 50 ng mL<sup>-1</sup> RANKL and 25 ng mL<sup>-1</sup> M-CSF to differentiate into osteoclast cells. On the other hand, osteoblast assay, 50μg mL<sup>-1</sup> ascorbic acid and 10 mM β-glycerophosphate were added to support differentiation. For control, the same cells were used without supplementation of respective growth factors. Biochemical analysis for osteoblast, i.e., Alkaline Phosphatase (ALP) activity was determined on day 3-14. The results showed that the ALP activity in the differentiation medium is significantly increased (p<0.05) on day 10 and day 14 as compared to control, i.e., cells without growth factors. Tartrate Resistant Acid Phosphatase (TRAP) activity that represented as osteoclast biomarker also showed significant increased (p<0.05) from day 3 until day 10 in the present of RANKL and M-CSF. The viability of differentiated cells also showed that the cells were able to survive until 10 to 14 days in the presence of respective growth factors without significant increased in respective differentiation medium. RT-PCR analysis on isolated RNA from mononucleated cells after 14 and 10 days in their differentiation medium showed that the osteoblast and osteoclast were expressing ALP (~373 bp) and TRAP (~281 bp) gene, respectively. Mononucleated cells originated from peripheral blood have the potential to differentiate into osteoblast and osteoclast cells in the presence of specific growth factors. The respective cells are primitive enough to differentiate into two distinct types of mature cells hence can be categorized as multipotent stem cells.
    No preview · Article · Mar 2008 · Journal of Biological Sciences