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

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.

Introducing Mammalian Cell Colony Formation in the Undergraduate Biology Laboratory †

“…Instructor preparation. Detailed information on how instructors can best prepare to teach and implement using HeLa cells in the undergraduate lab can be found in our previous publication (5).…”

A Thi2p Regulatory Network Controls the Post-glucose Effect of Xylose Utilization in Saccharomyces cerevisiae

“…Yeast cells were harvested at 20, 36, and 48 h, and diluted to a suitable multiple (~ 6 ×10 7 cells mL −1 ). The cells were then incubated with 0.04% trypan blue for 3–10 min; dead cells were stained by trypan, while live cells were not (Bowey-Dellinger et al, 2017). The stained cells were counted manually using a hemocytometer.…”

In vitro Evaluation of Seaweed Gracilaria verrucosa for Cytotoxic Activity against Cervical HeLa Cells

“…This is done to avoid inaccuracy in the results of the assay. 9 The passage cell used here will be passage 5 cell obtained from Department of Pathology Anatomy Faculty of Medicine University of Indonesia. The next step is to input the cultured cells into 96-microwell with each well containing 10 4 cells.…”

“…Absorbance is measured in 590 nm on a microplate reader (Model 550, Bio-Rad, USA). [9][10] Inhibitory activity of the extracts are measured using the following formula. Table 3.…”