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- 저자 : ( Chua Kien Hui ) (검색결과 3 건)
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Chua Kien Hui,Wan Kamarul Zaman Wan Safwani,Seah Shiao Chin,Annisaa Abu Samah Abdul Malek,Noormazita Hassan,Muhamad Syakeer Fazil,Raja Abdul Wafy Raja Muhammad Rooshdi,Adila A. Hamid,Somasundaram Sath 한국생물공학회 2012 Biotechnology and Bioprocess Engineering Vol.17 No.6
Recently human adipose-derived stem cells (ASCs) have shown much therapeutic potential in regenerative medicine. However, fetal bovine serum (FBS)used in culturing human cells may give risk to viral and prion transmission as well as immune rejection. Human serum (HS) is a safer growth supplement in human cell culture but its effects have not been well established. Therefore the objectives of this study were to compare the effects of HS versus FBS on the proliferation and stemness gene expression of ASCs. ASCs were cultured for 5passages in medium supplemented with either 10% HS or 10% FBS. ASCs proliferation rate and viability were determined at every passage. Total RNA was extracted at passage 5 (P5) and quantitative PCR was carried out to determine the stemness gene expression level of SOX-2,Nanog3, BST-1, REX-1, ABCG2 and FGF-4. The results showed ASC cultured in 10% HS scored greater proliferation rates and viability compared to 10% FBS. ASCs proliferated significantly faster in 10% HS compared to 10% FBS at P2, P3, and P4 (p < 0.05). In quantitative gene expression analysis, ASCs cultured in 10% FBS showed a significant increase of BST-1, REX-1 and ABCG2 expression compared to 10% HS. In conclusion, HS promotes ASCs proliferation and viability but its ability to support the stemness property of ASCs was inferior to FBS.
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( Norzana Abd Ghafar ),( Ropilah Abd Rahman ),( Jemaima Che Hamzah ),( Chua Kien Hui ),( Fauziah Othman ),( Aminuddin Bs3 ),( Ruszymah Bhi ) 한국조직공학·재생의학회 2007 조직공학과 재생의학 Vol.4 No.4
This study was designed to investigate the feasibility of using a serum-free and feeder layer-free culture system on serial expansion of rabbit limbal and corneal epithelial cells by evaluating the cell morphology, growth kinetics and gene expression pattern. Isolated epithelial cells(EC) from limbal and corneal regions of the rabbit corneoscleral tissues were culture expanded until three passages following initial culture in serum-free medium(SFM) supplemented with human corneal growth supplement(HCGS). EC morphology was examined daily with cell count and viability recorded at each passage by trypan blue exclusion test. Growth rate, number of cell doublings and total cell yield were measured and compared in each passage. The expression of Integrin β1; putative epidermal stem cell marker; Cytokeratin 12 and Connexin 43; differentiation markers for corneal epithelium; were detected by semiquantitative reverse transcriptase-polymerase chain reaction(RT-PCR). Limbal EC was able to retain compact and small cells morphology throughout the culture period. Corneal EC demonstrated a varying morphology with abundance of large, squamous cells and showed senescence earlier in culture. Limbal EC exhibited a greater proliferative activity demonstrated by higher growth rate, cumulative cell doublings and total cell yield compared to corneal EC. Integrin β1 expression was detected in freshly isolated and cultured limbal EC in all passages with relatively strong expression noted at the initial passage (P0). Corneal EC showed a very weak expression of Integrin β1 throughout the culture period. Cytokeratin 12 and connexin 43 were expressed in both freshly isolated limbal and corneal EC. Both corneal differentiation markers were expressed in cultured limbal and corneal EC in serial passages. These results suggested that corneal epithelium can be serially expanded in serum-free and feeder layer-free culture system with limbal EC showing a greater proliferative capacity while retaining its stemness via putative epidermal stem cell marker; Integrin β1 expression.
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( Munirah Sha`ban ),( Aminuddin Bin Saim ),( Samsudin Osman Cassim ),( Chua Kien Hui ),( Fuzina Nor Hussein ),( Ruszymah Bt Hj Idrus ) 한국조직공학과 재생의학회 2005 조직공학과 재생의학 Vol.2 No.4
This study was designed to verify the optimal · basic culture media that promote chondrocytes proliferation in vitro in order to facilitate adequate amount of chondrocytes for cartilage reconstructionas well as maintaining cartilage specific phenotype. Human articular chondrocytes were cultured in three types of basic culture media Ham`s F12, DMEM and the equivalent mixture of F12:DMEM. Cultured chondrocytes were trypsinized as they reached confluency. The viability and total number of cell were recorded at every passage. Large-scale culture expansion was used to reconstruct tissue-engineered cartilage. Quantitative RT-PCR analysis was used to evaluate the expression of collagen Type II, collagen Type I, aggrecan and Sox 9 gene, both in monolayer culture and in the engineered cartilage. The mixture of F12:DMEM promotes significantly greater (p<0.05) chondrocytes proliferation at every passage compared to the individual medium. Monolayer cultured chondrocytes exhibited down-regulation expression pattern of collagen Type II gene, aggrecan and Sox 9, whilst the expression of collagen Type I is up-regulated. Tissue-engineered cartilage morphologically and histologically resembled normal hyaline cartilage. Moreover, tissue-engineered cartilage re-expressed the specific chondrogenesis markers; collagen Type II, aggrecan and Sox 9. In conclusion, the mixture of F12:DMEM enhanced human articular chondrocytes proliferation thus provided adequate amount of chondrocytes for cartilage reconstruction. The new cartilage formed phenotypically resembles native cartilage. This results hold promise for the use of tissue-engineered cartilage implant for future orthopaedic reconstructive surgery.
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