Cell and Tissue Culture Foundations
Instructor: Scott Warila, Community College of Rhode Island
Location: CCRI
Session 1: June 23-24, June 26
Session 2: July 14-15, July 17
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.
Proposed Timeline (subject to change)
| Monday | Wednesday | Friday | |
|---|---|---|---|
| 9:00 AM-10:00 AM | Introduction to course/Overview of bacterial cell culture and bacterial transformation (lecture) | Intro to Bioreactors (lecture) Introduction to the 2L bioreactor and preparation for sterilization (Demo and hands- on) | Observation of E. coli liquid cultures from Day 2 Inoculation and operation of the 2L bioreactor (demo and hands-on) |
| 10:00 AM-11:00 AM | Preparing LB agar for selection of transformed E. coli (Demo and hands-on) | Calibration of 2L bioreactor DO and pH probes and sterilization of the bioreactor by autoclave (hands-on) | Observation of student prepared CHO cells from Days 1 and 2. Cryopreservation of CHO cells (hands-on) |
| 11:00 AM-12:00 PM | Transformation of E. coli with GFP plasmid (hands-on) | Examination of LB agar plates from Day 1. Selection of GFP producing colonies to inoculate LB broth (hands-on) | STEM cell culture with focus on myocardiocytes and tissue regeneration (discussion and demo) |
| 12:00 - 1:00 PM | Break | Break | Closing remarks WDT Survey Certificate Distribution |
| 1:00 PM-2:00 PM | Introduction to the CO2 incubator, BSC, aseptic technique, and preparing CHO cell media (hands-on) | Examination of instructor prepared CHO cell cultures using Floid cell imager. Perform a viable cell count and glucose/lactate assay. | |
| 2:00 PM-3:00 PM | Inoculating CHO cell media in T-flasks and plates from cryovial and culturing in CO2 incubator (hands-on) | Perform a 3-1 split of >70% confluent T-flasks (hands-on) | |
| 3:00 PM-4:00 PM | Wrap-up, review, and looking ahead (discussion) | Wrap-up, review, and looking ahead (discussion) |