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Short-term Course in Human Embryonic Stem Cell Culture Techniques

A 5-day course in human embryonic stem cell (hESC) culture techniques, which will cover the following topics:

  1. Introduction: brief scientific background, hESC markers
  2. Feeder cells
    • Derivation, culture, freezing and thawing of human and mouse feeder cells
    • Preparation of feeder-layer-covered, ready-to-use plates
  3. hESC culturing methods
    • Demonstration of hESC morphology
    • Culture and daily care
    • Freezing and thawing
    • Various culture media
  4. hESC differentiation methods
    • Embryoid body (EB) formation

Description of course topics

  1. Introduction. The introduction will include scientific background, general information on the course and safety precautions.
  2. Demonstrations. The demonstrations will include examples of hESC, feeder layer cultures, and EB formation.
  3. Practice. A hands-on personal experience where each attendant follows the protocols introduced during the course. The attendants will be given hESCs and feeder layer cultures to maintain during the course.
  4. Mouse embryonic fibroblast (MEF) culturing. Focus on MEF cultures with emphasis on MEF quality, rate of growth, viability and post thawing/splitting recovery.
  5. hESC culturing. Focus on hESC cultures, highlighting the expected rate of growth, viability, differentiated and undifferentiated morphology and post thawing or splitting recovery. Special attention will be paid to the morphology of differentiated and undifferentiated cells due to the difficulty involved in distinguishing between the two.
  6. EBs. Normal EB development. EBs at different stages of development will be prepared in advance in order to be examined by the participants.

    During the course, hESC, MEF and EB culture maintenance will be practiced, including daily medium preparation, medium exchange and splitting as required.

  7. Foreskin feeders. Demonstration of different stages in the derivation (if possible) and culture of foreskin lines.
  8. Guided individual practice. Attendants will be allocated time to repeat and practice the protocols as an opportunity to improve their performance. The instructors will be present for additional guidance and troubleshooting.
  9. Concluding remarks and discussion. An open discussion will be held at the end of the course, which will review and summarize the main points learnt. Attendants will be encouraged to take part in the discussion in the way of asking questions and highlighting any difficulties encountered during the course.
Course Timetable
Day 1 Day 2 Day 3 Day 4 Day 5
8:00–9:00 Reception, refreshments Reception, refreshments Reception, refreshments Reception, refreshments Reception, refreshments
09:00–10:00 Receiving cells Early embryo development Derivation of hESCs and culture methods: feeder free and human feeders.

comparison of different human feeders—Dr. Linzhao Cheng

 Follow up on hESCs and MEFs
10:00–11:00 Demondstration of hESC thawing ESCs specific markers hESCs follow up MEF freezing practice
11:00–12:00 Thawing hESCs -practice. Gene expression, Arrays MEFs follow up
12:00–13:00 Lunch break Lunch break Lunch break Lunch break Lunch break
13:00–14:00 MEF/foreskin splitting Practice MEF derivation Demonstration of hESC splitting Demonstration of EB generation and practice Open discussion and summary
14:00–15:00 Production of MEF/foreskin-covered plates Practice—hESC splitting MEF freezing demonstration Explanation on follow-up communication
15:00–16:00 MEF/foreskin thawing practice Demonstration and practice—hESC freezing MEF freezing practice Ending remarks and close of course
16:00–17:00 Discussion on daily practice Discussion on daily practice Discussion on daily practice Discussion on daily practice Social hour lunch or dinner

Location

Johns Hopkins University
Homewood Campus
Clark Hall, Building 35
Baltimore. MD, USA

Download the registration form for the January 12–16, 2009, offering (188k PDF; Get Adobe Reader)

Contact for further information:

Joseph Itskovitz-Eldor, M.D. D.Sc.
Director, Dept. of Obstetrics & Gynecology
Rambam Medical Center & The Faculty of Medicine, Technion
POB 9602, Haifa 31096
Israel

Email: Itskovitz@rambam.health.gov.il