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Cell Culture Techniques

Cell Culture Techniques. 程洪强，方瑜，李艳伟 2015 年 1 月. Cell Thawing and Migration Assay. Cell Thawing. Transfer cryovials containing frozen cells from -80℃ to a 37 ℃ water bath. Hold the cryovial on the surface of the water bath with an occasional gentle “flick” during thawing.

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Cell Culture Techniques

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  1. Cell Culture Techniques 程洪强，方瑜，李艳伟 2015年1月

  2. Cell Thawing and Migration Assay

  3. Cell Thawing • Transfer cryovials containing frozen cells from -80℃ to a 37 ℃ water bath. • Hold the cryovial on the surface of the water bath with an occasional gentle “flick” during thawing. • Dry off the outside of the cryovials and wipe with a 70% ethanol solution before opening the vial to prevent contamination. • Transfer the contents of one vial of cells to a 100 mm plate containing 10 ml of medium. • Incubate the cells at 37 ℃ /5% CO2, take a picture 2 hrs later

  4. Wound Assay • Before assay, cells are confluent in 6 well plates • Rinse with PBS and starvation in 0.5% serum overnight • Draw a line and mark on the bottom of the plate • Using a sterile 200 ul tip, scratch a wound through the cells • Rinse the cells gently with PBS and replace with medias (with or without serum) • Take pictures at 0, 24 and 48hr, measure the width, figure

  5. Marker and Scratch Your name, date, cell name Mark line scratch

  6. Arrangement of the scratches W/O serum W serum

  7. A representative result B - + TGFβ1 0 h 16 h 40 h

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