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Introducing Mammalian Cell Culture and Cell Viability Techniques in the Undergraduate Biology Laboratory

    Authors: Kristen Bowey-Dellinger1,2, Luke Dixon1, Kristin Ackerman1, Cynthia Vigueira1, Yewseok K. Suh1, Todd Lyda1, Kelli Sapp1, Michael Grider1, Dinene Crater1, Travis Russell1, Michael Elias2, V. McNeil Coffield1,*, Verónica A. Segarra1,*
    VIEW AFFILIATIONS HIDE AFFILIATIONS
    Affiliations: 1: Department of Biology, High Point University, High Point, NC 27268; 2: Department of Nanoscience, Joint School of Nanoscience and Nanoengineering, University of North Carolina at Greensboro, Greensboro, NC 27401
    AUTHOR AND ARTICLE INFORMATION AUTHOR AND ARTICLE INFORMATION
    • Received 17 November 2016 Accepted 16 April 2017 Published 11 August 2017
    • ©2017 Author(s). Published by the American Society for Microbiology.
    • [open-access] This is an Open Access article distributed under the terms of the Creative Commons Attribution-Noncommercial-NoDerivatives 4.0 International license (https://creativecommons.org/licenses/by-nc-nd/4.0/ and https://creativecommons.org/licenses/by-nc-nd/4.0/legalcode), which grants the public the nonexclusive right to copy, distribute, or display the published work.

    • Supplemental materials available at http://asmscience.org/jmbe
    • *Corresponding authors. Mailing address: V. McNeil Coffield: One University Parkway, High Point, NC 27268 Phone: 336-841-9394. E-mail: vcoffiel@highpoint.edu. Verónica A. Segarra: One University Parkway, High Point, NC 27268 Phone: 336-841-9507. E-mail: vsegarra@highpoint.edu.
    Source: J. Microbiol. Biol. Educ. August 2017 vol. 18 no. 2 doi:10.1128/jmbe.v18i2.1264
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    Abstract:

    Undergraduate students learn about mammalian cell culture applications in introductory biology courses. However, laboratory modules are rarely designed to provide hands-on experience with mammalian cells or teach cell culture techniques, such as trypsinization and cell counting. Students are more likely to learn about cell culture using bacteria or yeast, as they are typically easier to grow, culture, and manipulate given the equipment, tools, and environment of most undergraduate biology laboratories. In contrast, the utilization of mammalian cells requires a dedicated biological safety cabinet and rigorous antiseptic techniques. For this reason, we have devised a laboratory module and method herein that familiarizes students with common cell culture procedures, without the use of a sterile hood or large cell culture facility. Students design and perform a time-efficient inquiry-based cell viability experiment using HeLa cells and tools that are readily available in an undergraduate biology laboratory. Students will become familiar with common techniques such as trypsinizing cells, cell counting with a hemocytometer, performing serial dilutions, and determining cell viability using trypan blue dye. Additionally, students will work with graphing software to analyze their data and think critically about the mechanism of death on a cellular level. Two different adaptations of this inquiry-based lab are presented—one for non-biology majors and one for biology majors. Overall, these laboratories aim to expose students to mammalian cell culture and basic techniques and help them to conceptualize their application in scientific research.

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References & Citations

1. Rodríguez-Hernández CO, Torres-Garcia SE, Olvera-Sandoval C, Ramirez-Castillo FY, Muro AL, Avelar-Gonzalez FJ2014Cell culture: history, development and prospectsInt J Curr Res Acad Rev2188200
2. del Carpio A2014The good, the bad, and the HeLaBerkley Sci RevSpringhttp://berkeleysciencereview.com/article/good-bad-hela/
3. Masters JR2002HeLa cells 50 years on: the good, the bad and the uglyNat Rev Cancer231531910.1038/nrc77512001993 http://dx.doi.org/10.1038/nrc775
4. Schwarz E, Freese UK, Gissmann L, Mayer W, Roggenbuck B, Stremlau A, zur Hausen H1985Structure and transcription of human papillomavirus sequences in cervical carcinoma cellsNature31411111410.1038/314111a02983228 http://dx.doi.org/10.1038/314111a0
5. Skloot R2010The immortal life of Henrietta LacksRandom HouseNew York, NY
6. Emmert E2013Biosafety guidelines for handling microorganisms in the teaching laboratory: development and rationaleJ Microbiol Biol Educ141788310.1128/jmbe.v14i1.531238583563706168 http://dx.doi.org/10.1128/jmbe.v14i1.531
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2017-08-11
2017-11-21

Abstract:

Undergraduate students learn about mammalian cell culture applications in introductory biology courses. However, laboratory modules are rarely designed to provide hands-on experience with mammalian cells or teach cell culture techniques, such as trypsinization and cell counting. Students are more likely to learn about cell culture using bacteria or yeast, as they are typically easier to grow, culture, and manipulate given the equipment, tools, and environment of most undergraduate biology laboratories. In contrast, the utilization of mammalian cells requires a dedicated biological safety cabinet and rigorous antiseptic techniques. For this reason, we have devised a laboratory module and method herein that familiarizes students with common cell culture procedures, without the use of a sterile hood or large cell culture facility. Students design and perform a time-efficient inquiry-based cell viability experiment using HeLa cells and tools that are readily available in an undergraduate biology laboratory. Students will become familiar with common techniques such as trypsinizing cells, cell counting with a hemocytometer, performing serial dilutions, and determining cell viability using trypan blue dye. Additionally, students will work with graphing software to analyze their data and think critically about the mechanism of death on a cellular level. Two different adaptations of this inquiry-based lab are presented—one for non-biology majors and one for biology majors. Overall, these laboratories aim to expose students to mammalian cell culture and basic techniques and help them to conceptualize their application in scientific research.

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Figures

Image of FIGURE 1

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FIGURE 1

A schematic outlining the experimental procedure to count cells. PBS = phosphate-buffered saline; FBS = fetal bovine serum.

Source: J. Microbiol. Biol. Educ. August 2017 vol. 18 no. 2 doi:10.1128/jmbe.v18i2.1264
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Image of FIGURE 2

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FIGURE 2

Data from non-biology majors representing HeLa cell viability based on temperature. Student viability counts are shown from the incubation of HeLa cells at 4°C, 37°C, and 42°C. A) Representative data from a student pair that graphed average cell counts. Standard deviation is indicated as error bars where =3 for all treatments. B) Representative data from a student pair that graphed individual cell counts for each treatment.

Source: J. Microbiol. Biol. Educ. August 2017 vol. 18 no. 2 doi:10.1128/jmbe.v18i2.1264
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Image of FIGURE 4

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FIGURE 4

Student scores on lab deliverables suggest learning objectives were met. Student lab graphs were graded by instructors using the assessment rubric described above ( Table 2 ). The values shown correspond to students taught in the fall of 2016 ( > 100). A) Student average scores for both Biology majors and nonmajors. Standard deviation is indicated as error bars. B) Student scores as a percentage of students attaining a failing, passing, or high passing score.

Source: J. Microbiol. Biol. Educ. August 2017 vol. 18 no. 2 doi:10.1128/jmbe.v18i2.1264
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Image of FIGURE 3

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FIGURE 3

Data from biology majors representing HeLa cell viability based on diverse treatments. A) Student viability data from the incubation of HeLa cells with ethanol concentrations ranging from 0 to 25%. Standard deviation is indicated as error bars where =3 for all treatments. B) Student viability data from the incubation of HeLa cells with NaCl concentrations ranging from 0 to 0.5 M. Standard deviation is indicated as error bars where =2 for all treatments.

Source: J. Microbiol. Biol. Educ. August 2017 vol. 18 no. 2 doi:10.1128/jmbe.v18i2.1264
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