Cell and Tissue Culture Foundations
Instructor: Scott Warila, Community College of Rhode Island
Location: CCRI
Session 1: June 10, June 12, and June 14
Session 2: June 17, June 19, and June 21
Course Overview
This WDT module is designed to teach students the concepts of cell and tissue culture using bacterial and mammalian cells. Students will receive instruction in cell and tissue culture theory and perform hands-on activities to grow and maintain cell lines. This will include learning aseptic technique, working in a biological safety cabinet (BSC), preparing sterile media, using an autoclave, and inoculating cultures. The cultures will be grown in shake flasks, T-flasks, and a 2L bioreactor. The module will be completed over 2.5 days on a Monday, Wednesday, Friday schedule. Participants who complete the module will receive a RI-INBRE Certificate of completion.
Learning Outcomes
- Understand the concept of tissue culture
- Understand how a bioreactor operates
- Understand how an autoclave operates
- Learn how to prepare sterile media
- Learn how to inoculate a cell culture
- Demonstrate aseptic technique
- Become proficient working in a BSC
- Become proficient working with shake flasks and T-flasks
Electronic Laboratory Notebook
Students will be expected to maintain an electronic laboratory notebook in the cohort’s Google classroom. Each student will be given a generic lab template to complete, and students should record an entry for each procedure containing the title of the procedure, date performed and lab partner if applicable. They should write a brief description of the procedure and materials required. They should also record critical steps performed, amounts of reagents used, calculations, and any observations made. Lastly, conclusions should discuss steps that went well and steps that did not go well, and ways that the procedure could be improved to avoid mistakes in the future.
Additional Information:
It is recommended that students bring a tablet, iPad, or laptop to make entries in their electronic lab notebook. Lecture slides and laboratory handouts will be provided to students.
Timeline
| Monday | Wednesday | Friday | |
|---|---|---|---|
| 9:00 AM-10:00 AM | Introduction to course/Overview of bacterial cell culture (Lecture) | Intro to Bioreactors (lecture) | Observation of E. coli liquid cultures from Day 2 (hands-on) |
| 10:00 AM-11:00 AM | Bacterial transformation and the GFP plasmid (lecture) Setting up and using electronic notebooks (demo and hands-on) | Calibration of 2L bioreactor DO and pH probes and sterilization of the bioreactor by autoclave (hands-on) | Inoculation and operation of the 2L bioreactor (demo and hands-on) |
| 11:00 AM-12:00 PM | Preparing LB media (agar and broth) for selection of transformed E. coli (Demo and hands-on) | Examination of LB agar plates from Day 1. Selection of GFP producing colonies to inoculate LB broth (hands-on) | Observation of CHO cells from Day 2. Perform a viable cell count. Glucose/lactate assay and 2-1 split of >70% confluent T-flasks (hands-on) |
| 12:00 - 1:00 PM | Break | Break | Closing remarks WDT Survey Certificate Distribution |
| 1:00 PM-2:00 PM | Transformation of E. coli with GFP plasmid (hands-on) | Introduction to the CO2 incubator, BSC, aseptic technique, and preparing CHO cell media (hands-on) | |
| 2:00 PM-3:00 PM | Introduction to the 2L bioreactor and preparation for sterilization (Demo and hands-on) | Inoculating CHO cell media in T-flasks and culturing in CO2 incubator (hands-on) | |
| 3:00 PM-4:00 PM | Wrap-up, review, and looking ahead (discussion) | Wrap-up, review, and looking ahead (discussion) |