Cell Monitoring and Remote Recording: The CytoSMART System

Transcription

1 Pharma&Biotech BioResearch Cell Monitoring and Remote Recording: The CytoSMART System Webinar 22 September 2015 / Speaker: Sean Fuerst 23 September 2015 / Speaker: Dr. Claudia Schwartz Lonza Cologne GmbH, Cologne / 2015 Lonza

2 Key Observations for Precious Cell Cultures In general: Cell growth and confluency Time to passage Tracking of detaching process Cell attachment Quality of culture and cell death For ipsc culture: Spontaneous differentiation Reprogramming Directed differentiation and morphology change 2

3 Agenda Introduction to the CytoSMART System Cell culture standardization Applications for ipsc culture Remote monitoring of precious cell cultures The CytoSMART System How it works CytoSMART Connect Cloud Project Page Summary 3

4 Agenda Introduction to the CytoSMART System Cell culture standardization Applications for ipsc culture Remote monitoring of precious cell cultures The CytoSMART System How it works CytoSMART Connect Cloud Project Page Summary 4

5 CytoSMART System Components + CytoSMART Device With 10X objective Tablet with CytoSMART App CytoSMART Connect Cloud 5

6 CytoSMART System How Does it Work? 6

7 CytoSMART System: Time Lapse Video View the actual video in the webinar recording: 7

8 CytoSMART System Main Features Easy handling Set-up of system within minutes Small size Easily fits in every incubator Cloud technology Monitor your cell culture anywhere and anytime Low cost The affordable live cell imaging tool ideal for small budgets 8

9 Agenda Introduction to the CytoSMART System Cell culture standardization Applications for ipsc culture Remote monitoring of precious cell cultures The CytoSMART System How it works CytoSMART Connect Cloud Project Page Summary 9

10 Importance of Cell Culture Standardization Condition of a cell culture has significant influence on down stream results Cell interactions play an important role in cell behavior Standardize cell culture by monitoring cell confluency 10

11 Effect of Confluency on Transfection Efficiency Neonatal Human Epidermal Keratinocytes (NHEKneo) were cultured until they reached 40 and 70% confluency. Cells were transfected with pmaxgfp Vector using the Nucleofector Technology under the same conditions. Transfection efficiencies were determined by flow cytometry. 100 CytoSMART Images: % confluency Transfection efficiency [%] % 70% 70% confluency Confluency 11

12 Impact of Cell Confluency on Growth Behaviour AoSMC cultured to 70% and 100% confluency, passaged and seeded (at 8x10 4 cells/cm 2 ) Proliferation monitored for 3.5 days Growth curve determined by CytoSMART App CytoSMART Images: Confluency [%] Time [hours] 70% confluency Time [hours] 100% confluency 12

13 Monitoring of Cell Confluency The CytoSMART System takes images of the cell culture at predefined time points (every 1 to 60 min) An algorithm determines cell confluency of the cell monolayer at all time points Growth curve is displayed 13

14 Automatic Confluency Alert Automatic confluency alerts can be requested on project page in Notifications Pane Percentage of confluency can be individually selected between 1 and 100% Automatic alerts are sent to your address 14

15 Summary: CytoSMART System for Cell Culture Standardization Standardization of cell culture is important as the condition of a cell culture has significant influence on down stream results Automatic alerts help to standardize cell culture Document growth curves of your cell culture, e.g. for new cell cultures Screen effects of various factors, such as culture media and growth factors, on your cell culture 15

16 Agenda Introduction to the CytoSMART System Cell culture standardization Applications for ipsc culture Remote monitoring of precious cell cultures The CytoSMART System How it works CytoSMART Connect Cloud Project Page Summary 16

17 Watch and Document Your ipsc Cell Culture ipsc culture is different from (primary) cell culture Culture properties to be monitored Cell attachment Spontaneous differentiation Cell growth and confluency Quality of culture and cell death Time to feed / passage Reprogramming Directed differentiation and morphology change 17

18 ipsc Culture for Beginners How the CytoSMART System Can Help Time point of subculture Tracking of passaging process Spontaneous differentiation of ipscs 18

19 PSC Applications: Cell Culture Media Evaluation L7 XF/EOD View the actual video in the webinar recording: Video of H7 hescs P60 taken with CytoSMART System for 3 days (72 hours) 19

20 PSC Applications: Cell Culture Media Evaluation Competitor View the actual video in the webinar recording: Video of H7 hescs at P60 taken with CytoSMART System for 2 days (48 hours) 20

21 ipsc Applications: Spontaneous Differentiation (No Feeding in 3 Days) 0 h 8 h View the actual video in the webinar recording: Video of hipscs (ENG 5 B P23) taken with CytoSMART System for 2 days (48 hours) 21

22 ipsc Application: Colony Growth (Details) View the actual video in the webinar recording: 0 h 8 h Video of hipscs (ENG 5B P23) taken with CytoSMART System for 52h 22

23 Agenda Introduction to the CytoSMART System Cell culture standardization Applications for ipsc culture Remote monitoring of precious cell cultures The CytoSMART System How it works CytoSMART Connect Cloud Project Page Summary 23

24 Where Can the CytoSMART System Assist to Monitor Precious Cell Cultures? First example: Culturing cells under physiological conditions in a hypoxia workstation Working in closed systems: necessary to keep O 2 and CO 2 content, temperature and humidity constant Cell monitoring via classical microscope is cumbersome and interrupts constant culture under hypoxia Solution: Let the CytoSMART System take images of the cell culture and view the cells remotely Pictures of hypoxia workstations kindly provided by BioSpherix (left), Oxford Optronix (center) and HypOxygen (right) 24

25 Where Can the CytoSMART System Assist to Monitor Precious Cell Cultures? Second example: Culturing cells in a GMP environment In GMP suite, even observation of the culture requires trained staff and significant labor time 25

26 Agenda Introduction to the CytoSMART System Cell culture standardization Applications for ipsc culture Remote monitoring of precious cell cultures The CytoSMART System How it works CytoSMART Connect Cloud Project Page Summary 26

27 How It Works Place cell culture vessel with CytoSMART Device in incubator and connect to tablet Follow the intuitive CytoSMART App on the tablet View cell culture remotely via CytoSMART Connect Cloud from your computer, tablet or smartphone How-it-works video on 27

28 CytoSMART Connect Cloud Project Page Project summary Set automatic alerts for cell coverage and temperature Notes field (optional entry) Download functionality (.jpeg/.avi/.csv). Adjust speed of movie View your cell culture Press start to watch video of your cell culture Cell coverage graph Move red line to view your cells in big image at chosen time Temperature graph 28

29 CytoSMART System Summary of Applications Cell culture monitoring Cell culture standardization Automatic alerts for confluency, recording of cell growth curves and screening of culture media and growth factors Stem cell culture documentation Remote monitoring under hypoxic cell culture and GMP conditions or of ipsc reprogramming Migration assays Automatic photo/video documentation (instead of manual process) 29

30 Summary Easy handling Small size Cloud technology Low cost 30

31 Become CytoSMART Lab of the Month Register online at to receive a CytoSMART System for one month for free Benefit from the CytoSMART System to monitor and record your preferred cell culture We promote your research activities and share your favorite time lapse recording with the research community on our website 31

32 Contact Details Cell Culture Experts Scientific Support Team US: or Scientific Support Team EU + ROW: scientific.support.eu@lonza.com or

33 Interested in Learning More? Join our upcoming webinar in October: Enhance Your Cell Culture with RAFT 3D Cell Culture System Register now: Sign up for our Cell Culture or Transfection enews Follow us on LinkedIn View or download our BioResearch Catalog 33

34 Thank You! All trademarks belong to Lonza or its affiliates or to their respective third party owners.