1 / 14

B IOMATERIALS L AB #3 Cell culture and cell-(bio)material Interactions

Explore cell culture techniques and cell-biomaterial interactions using PEGMA and Acr-PEG-RGDSG. Learn about linear molecules and safety protocols. Hands-on experiments include live/dead imaging and data collection with ImageJ.

bellomy
Download Presentation

B IOMATERIALS L AB #3 Cell culture and cell-(bio)material Interactions

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. BIOMATERIALS LAB #3Cell culture and cell-(bio)material Interactions Monday, February 27th and Tuesday, February 28th BME 245/445 – Lab 3

  2. PRE-LAB • Sign-in • Quiz (Benoit. 2005) BME 245/445 – Lab 3

  3. LINEAR MOLECULES • PEGMA • Acr-PEG-RGDSG Modified from (Lin, C.C., Pharmaceutical Research. 26(3): 631-643. 2009) BME 245/445 – Lab 3

  4. SECTION 1- Done before lab PEGMA Network Synthesis (done before lab – as previous) • 10wt% PEGMA in PBS • +/- 5 mM Acr-PEG-RGDSG • 10% photo-initiator by volume (LAP) • 0.75 mL Placed between two glass slides with spacer • Exposed to UV-lamp for 10min • Remove, punch out cylindrical samples • Place in PBS BME 245/445 – Lab 3

  5. SECTION 2 Cell Culture (TA demo and then hands on) • Sterile Techniques • Feeding cells • Counting cells • Seeding cells (TC plastic / PEG / PEG+RGDSG) • Monitor cell-material interactions over time… (ImageJ) • # of adherent cells • Average adhesion area • Average spindle factor BME 245/445 – Lab 3

  6. SECTION 3 Time-course data collection • Days 0, 1, 2, 3 • # adherent cells, adhesion area, spindle factor (ImageJ) • Day 2 • Live/Dead imaging BME 245/445 – Lab 3

  7. Lab Safety • BSL 1/2 • Human cell lines in culture room  BSL 2 • We’re using animal cells (Mouse embryonic fibroblasts)  BSL 1 • Hoods • Filter out biologic particulates • Gloves, glasses, caution • No food/drink/chapstick/ect

  8. STERILE TECHNIQUE • Gloves! • Spray everything down with EtOH before entering hood • Do NOT reach over the top of open containers • Do NOT leave containers open unless in use • Caps always placed right-side up • Change pipette tips after every use • Clean up after done • Soap for spills • EtOH to sterilize BME 245/445 – Lab 3

  9. IMAGEJ • Open ImageJ • File  Open  select image file • Area: Polygon option • Diameter: Line option • Analyze  Set Measurements (select area) • Analyze  Measure (area/length) BME 245/445 – Lab 3

  10. IMAGEJ Future measurements are calibrated Can add scale bars Draw line of known length Set global scale BME 245/445 – Lab 3

  11. IMAGEJ Line Selection (for Diameter) Polygon Selection (for Area) BME 245/445 – Lab 3

  12. SPINDLE FACTOR • Ratio of major axis to minor axis SF = 96.566/86.770 = 1.11 SF = 118.127/47.676 = 2.48 BME 245/445 – Lab 3

  13. BME 245/445 – Lab 3

  14. Scheduling BME 245/445 – Lab 3

More Related