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DC Field | Value | Language |
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dc.contributor.advisor | Muhammad Lokman Md. Isa, Ph.D | en_US |
dc.contributor.advisor | Mohd Hamzah Mohd Nasir, Ph.D | en_US |
dc.contributor.advisor | Afzan Mat Yusof, Ph.D | en_US |
dc.contributor.author | Jenuit, Melinda | en_US |
dc.date.accessioned | 2021-02-11T04:51:15Z | - |
dc.date.available | 2021-02-11T04:51:15Z | - |
dc.date.issued | 2020 | - |
dc.identifier.uri | http://studentrepo.iium.edu.my/handle/123456789/10261 | - |
dc.description.abstract | 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. | en_US |
dc.language.iso | en | en_US |
dc.publisher | Kuantan, Pahang : Kulliyyah of Science, International Islamic University Malaysia, 2020 | en_US |
dc.subject.lcsh | Fibroblast | en_US |
dc.subject.lcsh | Cells -- Growth -- Regulation | en_US |
dc.subject.lcsh | Animal cell biotechnology | en_US |
dc.subject.lcsh | Biodiversity conservation | en_US |
dc.title | Establishing a standard fibroblast cell line derived from kidney tissue of Sumatran rhinoceros (puntung) | en_US |
dc.type | Master Thesis | en_US |
dc.description.identity | t11100424165MelindaJenuit | en_US |
dc.description.identifier | Thesis : Establishing a standard a standard fibroblast cell line derived from kidney tissue of Sumatran rhinoceros /by Melinda Jenuit | en_US |
dc.description.kulliyah | Kulliyyah of Science | en_US |
dc.description.programme | Master of Science (Biotechnology) | en_US |
dc.description.abstractarabic | إن تَطْبِيقْ إِنْشَاءْ وَحِفْظْ الخَلَايَا بِالتَبْرِيدْ فِي التَنَوُعْ البَيُولُوجِي أَمْرٌ ضَرُورِي لِلْحَيَاةِ البَرِيّةِ المُهَدّدَةِ بِالِانْقِرَاضْ مِثِلْ وَحِيدُ القَرْنِ السُومَطْرِي. إِنّهَا بِمَثَابَةِ الأَسَاسْ لِتِقْنِيَاتْ خُلَوِيّة مُتَقَدِمَة. للحُصُولْ عَلَى الأَنْسِجَة المَرْغُوبَة مِنْ الكَائِنًاتِ الحَيّة كَمَصْدًرْ لِلْثَقَافَةِ الأَوّلِي غَيْرُ مُمْكِنْ. كَبَدِيلْ، تَمّ اِسْتِخْدَامْ أَنْسِجَة مَحْفُوظَةْ مِنْ وَحِيدْ القَرْنِ الذِيْ تَمّ قَتْلُهُ بِسَبَبِ سَرَطَانِ الجِلْدِ في مَرَاحِلِهِ الأَخِيرَة. الهَدَفْ مِنْ هَذِهِ الدِرَاسَة هُو تَحْدِيدْ وَتَمْيِيزْ وَتَوْثِيقْ الخَلاَيَا اللِيفِيّة المُشْتَقّة مِنْ أَنْسِجَةْ الكِلَى في جَسَدْ وَحِيدْ القَرْنْ السُومَطْرِي. تَمّ عَزْلْ الأَنْسِجَة الأَوَلِيةْ مِنْ أَنْسِجَةِ الكِلَى بِاِسْتِخْدَامِ طَرِيقَةْ زَرْعِ الإِنْزِيمْ المُخْتَلِطْ، تُسْتَكْمَلْ بَأَوْسَاطٍ كَامِلَة (وَسَطْ النَسَر المُعَدّلْ مِنْ قِبَلْ دُولْبِيكُو + 10% مَصِلْ بَقَرِي جَنِينِي + 1% مُضَادْ حَيَوِيْ) تم حْفَظْها عِنْدَ 37 دَرَجَة مِئَوِيَة مَعَ حَاضِنَه 5% مِنْ ثَانِي أُكْسِيْدِ الكَرْبُونْ. بَعْدَ التِرِبْسِينْ الرُوتِينِي، تَمّ الحُصُولْ عَلَى مُنْحَنَيَاتْ قَابِلِيّةْ الحَيَاةْ وَالنُمُو مِنْ خِلَالِ طَرِيقَةِ عَدّ زُرْقَةِ التِرْيبَانْ. تًمّ الاِحْتِفَاظْ بِالأَسْهُمْ المُجَمّدَة في وَسَاِئطْ تَحْتَوِي عَلَى (90% مِنْ مَصِلْ بًقَرِي جَنِينِي وَ10% مٍنْ ثُنًائِي مِيثِيلْ سُلُفْوكْسِيدْ) تَمّ تَحْدِيدْ الشَيْخُوخَةْ الخَلَوِيّة بِوَاسِطَةْ مُقَايَسَةْ تَلْطِيخْ إِنْ بِيتَا جَالَاكْتُوزِيدَازْ المُرْتَبِطْ بِالسِنِيسِنْسْ. بَيْنَمَا الكُرُومُوسُومَاتْ المُنْتَشِرَة مَصْبُوغَة بِمَحْلُولْ غِيْمْزَا. يُوَلّدْ اِشْتِقَاقْ خَلَايَا الأَرُوُمَة الِليفِيّة مِنْ أَنْسِجَة الكِلَى مَا مَجْمُوعُه 81 مِنْ الأَسْهُم المُجَمّدَة. حَافَظَتْ قَابِلِيّة الخَلِيّة عَلَى البَقَاءِ لِأَكْثَرِ مِنْ 80٪ خِلَالِ القَنَوَاتِ التَسَلْسُلِيّة وَبَعْد 3 أَشْهُرْ مِنْ الحِفِظْ بِالتَبْرِيدْ، وَلَكِنْ فَقَطْ الخَلايَا فِي ب5 وَ ب10 تُظْهِرْ شِفَاءً مَعْقُولًا بَعَدْ 6 أَشْهُرْ. مِنْ مُنْحَنَيَاتِ النُمُو، كَانَ عَدَدْ الخَلَايَا المُضَاعَفَة فِي ب5 هُوَ 20.45 سَاعَة بَيْنَمَا 22.35 سَاعَة فِي كُلّ مِنْ ب10 وَ ب15. يَزْدَادْ مُسْتَوَى الشَيْخُوخَة بِشَكِلْ مَلْحُوظْ مِنْ ب5 إلى ب10 ، وَخَاصَةً بِشَكْلٍ مَلْحُوظْ عِنْدَ ب15. اُعْتُبِرَتْ الثَبَاتَاتْ الجِينِيّة مُسْتَقِرّه عِنْدَ ب5 وَ ب10. كَانَتْ جَمِيعْ الثَقَافَاتْ خَالِيَة مِنْ تَلَوٌّثْ المِيكُوبَلاَزْمَا. | en_US |
dc.description.nationality | Malaysian | en_US |
dc.description.callnumber | t QP 88.23 J54E 2020 | en_US |
dc.description.notes | Thesis (MSBTS)--International Islamic University Malaysia, 2020. | en_US |
dc.description.physicaldescription | xiv, 89 leaves : colour illustrations ; 30cm. | en_US |
item.openairetype | Master Thesis | - |
item.grantfulltext | open | - |
item.fulltext | With Fulltext | - |
item.languageiso639-1 | en | - |
item.openairecristype | http://purl.org/coar/resource_type/c_18cf | - |
item.cerifentitytype | Publications | - |
Appears in Collections: | KOS Thesis |
Files in This Item:
File | Description | Size | Format | |
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t11100424165MelindaJenuit_24.pdf | 24 pages file | 856.76 kB | Adobe PDF | View/Open |
t11100424165MelindaJenuit_SEC.pdf Restricted Access | Full text secured file | 4.07 MB | Adobe PDF | View/Open Request a copy |
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