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== Test #1 (ASIC_E1_T1) == *Cells: osteogenic differentiated hESCs, 3 days after osteogenic induction(from H9_Bb2 P9) *Cell number: ~2.5*10e5/ml *Well number: A ===Plate operation:=== '''precoating''' 1. Prepare Matrigel (coating solution). 2. Aspirate solution from wells 1 and 8. Leave bottom holes fill with PBS. 3. Aspirate PBS from well 6 and 7, including bottom hole. 4. Add 10ul coating solution to well 6 (add to the bottom hole). 5. Incubate at room temperature for 1 hr. 6. Wash step: aspirate coating solution from well 6 and 7. Add 10ul culture medium to the bottom hole of well 6. '''Cell loading''' 1. Prepare cell suspension (Osteogenic differentiated hESCs from one 6-well plate, dilute 50X). 2. Aspirate solution from well 6 and 7, including bottom hole. 3. Pipette 10ul of cell suspension to well 6 (bottom hole). Monitor under microscope. Take pictures. '''Preculture''' 1. Aspirate solution from well 7, including bottom hole. 2. Add 350ul osteogenic induction medium to well 1, and 50ul medium to well 7. 3. Observe under microscope. Place plate in incubator. <br> ===Results:=== *Cell number was not enough. *Cells were clustered (close to inlet 6). Cell loading protocol need to be improved. *350ul culture medium could last for 3-4 days. *Some of the cells died after seeding, others expanded a bit and form colonies. No more expansion after 3 days. *Image after cell loading [[File:Tina Lo_021813_ASIC_E1_T1_pic 1.jpg]] [[File:Tina Lo_022113_ASIC_E1_T1_pic 2.jpg]] [[File:Tina Lo_022213_ASIC_E1_T1_pic 3.jpg]] <br> <br>
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