Cell culture assignement

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Carleton University *

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3332

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Biology

Date

Dec 6, 2023

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pdf

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4

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HSS 3109- Research Approaches in Health Biosciences Self-learning module on Cell culture For this short assignment, you will need to go to the ATCC website ( www.atcc.org ) to retrieve information that will allow you to answer the following questions: You have been hired by a small startup pharma that aims to develop pharmacological agents that can promote muscle cell differentiation. In order to test newly synthetized compounds the team asks you to propose a muscle cell culture model that uses normal i.e. non- diseased) cell lines that are commercially available through ATCC. The cell culture model should also be compatible with tools (antibodies and PCR primers) already available in house, which were developed in mouse. Before decisions are made they ask you to prepare a report that answers the following questions:
HSS 3109- Research Approaches in Health Biosciences 1. How many cell lines are derived from skeletal muscle irrespective of species? 10 cells lines are derived from skeletal muscle. 2. In the cell lines derived from skeletal muscle, how many are of myoblastic origin? There are 5 available cell lines that are from myoblastic origin. 3. How many myoblastic muscle cell lines are derived from normal tissue (is free of disease)? There is 4 myoblastic muscle cell lines derived from normal tissue. 4. Of those, how many are of mouse origin? 3 of the muscle cell lines are of mouse origin. 5. Based on the characteristics of these cell lines, which one would be the most appropriate? Based on the characteristics Sol8 will be the most appropriate cell line. 6. What is the origin of the Sol8 cell line (i.e. species)? The origin of Sol 8 cell line is from mus musculus. 7. What is the morphology and culture properties of the Sol8 cell line? Fibroblast is the morphology property and adherent are the culture property of the Sol8 cell line. 8. From which muscle was the Sol8 line derived from? It was derived from the skeletal muscle. 9. Are the Sol8 cells able to differentiate and if yes, under which conditions? Yes, they will differentiate into myotubes. The culture should be incubated at 37°C in a suitable incubator with 5% CO 2 in the air for best results. To maintain high viability, promptly thaw the vial and initiate the culture as soon as possible upon receipt. If further frozen storage is needed, it should be stored in liquid nitrogen vapor, not at -70°C. Storage at -70°C will result in loss of viability . 10. What is the cell culture media required? ATTC- formulated Dulbecco’s Modified Eagle’s Medium with fetal bovine serum to a final concentration of 20% 11. Are there any particularities related to sub-culturing for this cell line? One of the particularities are that it never allows cells to become confluent, and the sub-culture is at 50% of confluence. 12. How many media changes are required per week? Media changes are required 2 to 3 times per week 13. What is the recommended sub cultivation ratio? The recommended sub cultivation ratio is 1:18 to 1:22 14. What is the approximate population doubling time? 16 to 18 hours is the approximate population doubling time.
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