Establishing a standard fibroblast cell line derived from kidney tissue of Sumatran rhinoceros (puntung) /

Establishment and cryopreservation of cell cultures have been applied in biodiversity conservation, essential to critically endangered wildlife such as the Sumatran rhinoceros. It serves as the foundation for advanced cellular techniques. Obtaining desired tissues from living organism as a source fo...

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Bibliographic Details
Main Author: Jenuit, Mellinda (Author)
Format: Thesis
Language:English
Published: Kuantan, Pahang : Kulliyyah of Science, International Islamic University Malaysia, 2020
Subjects:
Online Access:http://studentrepo.iium.edu.my/handle/123456789/10261
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008 201013s2020 my a f m 000 0 eng d
040 |a UIAM  |b eng  |e rda 
041 |a eng 
050 0 0 |a QP88.23 
100 1 |a Jenuit, Mellinda,  |e author 
245 1 0 |a Establishing a standard fibroblast cell line derived from kidney tissue of Sumatran rhinoceros (puntung) /  |c by Melinda Jenuit 
264 1 |a Kuantan, Pahang :   |b Kulliyyah of Science, International Islamic University Malaysia,   |c 2020 
300 |a xiv, 89 leaves :  |b colour illustrations ;  |c 30cm. 
336 |2 rdacontent  |a text 
337 |2 rdamedia  |a unmediated 
338 |2 rdacarrier  |a volume 
338 |2 rdacarrier  |a online resource 
347 |2 rdaft  |a text file  |b PDF 
500 |a Abstracts in English and Arabic. 
500 |a "A thesis submitted in fulfilment of the requirement for the degree of Master of Science (Biotechnology)." --On title page. 
502 |a Thesis (MSBTS)--International Islamic University Malaysia, 2020. 
504 |a Includes bibliographical references (leaves 75-84). 
520 |a Establishment and cryopreservation of cell cultures have been applied in biodiversity conservation, essential to critically endangered wildlife such as the Sumatran rhinoceros. It serves as the foundation for advanced cellular techniques. Obtaining desired tissues from living organism as a source for primary culture is not feasible. As alternative, preserved tissues from Puntung which was euthanized due to terminal skin cancer was utilized. The aim of this study is to establish, characterize and authenticate fibroblast cells derived from kidney tissue of Sumatran rhinoceros carcass. Primary cultures were isolated from kidney tissue using mixed enzymatic-explant method, supplemented with complete media (DMEM + 10% FBS + 1% antibiotic) and kept at 37ºC with 5% CO2 incubator. Following routine trypsinization, viability and growth curves were obtained by Trypan blue counting method. Frozen stocks were preserved in media containing 90% FBS and 10% DMSO. Cellular senescence was quantified by Sa-β-gal staining assay, while chromosome spreads were stained with Giemsa solution. Mycoplasma contaminations were detected by PCR method. Derivation of fibroblast cells from the kidney tissues generates a total of 81 frozen stocks. The cell viability maintained over 80% during serial passages and after 3 months of cryopreservation, but only cells at P5 and P10 show reasonable recovery after 6 months. From the growth curves, the cell population doubling time at P5 was 20.45h while 22.35h at both P10 and P15. The senescence level significantly increase from P5 to P10, and especially significant at P15. Genetic stabilities were considered stable at P5 and P10. All cultures were free from mycoplasma contamination. The results of this study have concluded that cells cultured up to P10 were suitable for the development of Sumatran rhinoceros cell banking system, which are applicable for advanced research. This also provides simple reference in the development of cell bank for other endangered wildlife in Malaysia. 
596 |a 6 
650 0 |a Fibroblast 
650 0 |a Cells  |x Growth Regulation  
650 0 |a Animal cell biotechnology 
650 0 |a Biodiversity conservation 
655 7 |a Theses, IIUM local 
690 |a Dissertations, Academic  |x Department of Biotechnology  |z IIUM 
700 0 |a Muhammad Lokman Md. Isa,  |e degree supervisor 
700 0 |a Mohd Hamzah Mohd Nasir,  |e degree supervisor 
700 0 |a Afzan Mat Yusof,  |e degree supervisor 
710 2 |a International Islamic University Malaysia.  |b Department of Biotechnology 
856 4 |u http://studentrepo.iium.edu.my/handle/123456789/10261 
900 |a sz-asbh 
999 |c 440058  |d 473381 
952 |0 0  |6 T QP 000088.23 J54E 2020  |7 0  |8 THESES  |9 763372  |a KIMC  |b KIMC  |c CLOSEACCES  |g 0.00  |o t QP 88.23 J54E 2020  |p 11100424165  |r 1900-01-02  |t 1  |v 0.00  |y THESIS