Exp-5-ATC-Full.docx - PRACTICAL REPORT Marks Faculty...

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PRACTICAL REPORT Marks Faculty: Faculty of Science Course: Biomedical Science, Biotechnology, Agricultural Science Unit Code & Title : UDBA/UDAA2243 Animal Tissue Culture Trimester: January 2018 Title: Practical 5: Primary explants: Observation of primary cells from chick embryo fibroblast Practical Group: 2 Date of Submission: 30/3/2018 Instructor: Dr. Kwong Phek Jin Declaration : We certify that this report is our own work, where material and reference used from other sources have been properly acknowledged. Group members Name ID Number 1 MUHAMAD SYAMIL BIN ROSLI 1507809 2 CATHERINE SIEW LEE YENG 1507264 3 EUNICE MAH SHAO THEEN 1508143 4 KIMBERLY KUAN MIN YI 1506954 5 SOO CHI FOONG 1600761 6 MANDY YEN SOOK LENG 1507500 7 RAVEENDERAN SITHAMBARAM 1301399 1
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Introduction Cell culture is removal of cells from animal and their growth in a favorable artificial condition. The cells are disaggregated enzymatically or mechanically and taken directly from living organism. The cells are isolated from tissues and they are cultured under appropriate condition. Thus, they will proliferate until they occupy all substrate or reach confluency. When they reach 80-90% of confluency, they will be sub-cultured in which they will be transferred to a new flask with new medium. Thus, more substrates are available for them to proliferate or grow further. These cells are known as primary culture (ThermoFisher, 2018). The culture consists of mixed population of cell type. However, some of the cell type may not adhere to substrate or grow under appropriate condition. Several cell types have limited lifespan which is less than 5-20 cell division. Thus, the cell type balance change quickly with time in which the fast-growing cells outgrow the slow-growing cells or non-proliferating cells, causing loss of heterogeneity. The advantage of the loss of heterogeneity is that pure population can be obtained for development of cell lines if the predominant emerging cell types is the cell of interest. Thus, it is good for selection to produce a more homogeneous population. For disadvantage of selective growth, the heterogeneity and diversity of multicellular tumor is lost (Langdon, 2014). After subculture, the cells from explants which is primary cell culture become cell line or sub-clone. Cell lines from primary cell culture have a limited life span which is known as finite cell line. After sub-culture, cell populations with the highest growth rate will continue to proliferate for a number of cell generations, resulting in a uniform genotype and phenotype of population (Invitrogen, 2018). Objective 1. To learn the principle and technique in isolation of primary culture involving chick embryo fibroblast (CEF). 2. To observe the morphology and confluency of primary fibroblast cells from chicken embryo, colour change in media and contamination signs at different incubation period.
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