想用贴壁的方法培养神经干细胞？怎样包被圆盖片？

lhmxm

贴壁培养，怎么处理培养孔内的圆盖片？用的是什么粘附介质？包被之后是立即清洗，还是烘干后再清洗？

千细胞

可以考虑
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% F/ Q1 x& C& n$ j8 iPoly-L-ornithine-coated coverslips—In a sterile hood, put one sterilized coverslip in each well of a 24-well plate. Add 75 μl of 0.1 mg ml−1 poly-L-ornithine onto each coverslip. Incubate the plates at 37 °C overnight. The next day, aspirate the poly-L-ornithine and let the coverslips dry at roomtemperature for ~30 min. Wash three times using 1 ml of sterile water for each well. Leave the plate open in the hood until completely dry. Cover the plates, wrap in foil, label with date and store at −20 °C for up to 4 weeks.7 i( V) t$ p& ^% j4 E& m! V9 r
Laminin-coated six-well plate—Dilute laminin with fresh neural differentiation medium to a final concentration of 20 μg ml−1. Add 300 μl of laminin solution to each well of a sixwell plate. Allow the medium to hold as a big drop and spread within the central area of the well. Do not let the medium drain to the edge. Incubate the plate at 37 μC for 1 h. Laminin easily absorbs to plastic and tends to form aggregates at room temperature. Store laminin at −80 °C and thaw at 4 °C before using. Avoid aliquotting laminin into plastic tubes from the original glass vial.( a- [8 w! t& n. Z

千细胞

可以考虑% z. M! F$ I+ ?
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EQUIPMENT SETUP( U0 |  E2 c/ S5 o9 K# K
Poly-L-ornithine-coated coverslips—In a sterile hood, put one sterilized coverslip in each well of a 24-well plate. Add 75 μl of 0.1 mg ml−1 poly-L-ornithine onto each coverslip. Incubate the plates at 37 °C overnight. The next day, aspirate the poly-L-ornithine and let the coverslips dry at roomtemperature for ~30 min. Wash three times using 1 ml of sterile water for each well. Leave the plate open in the hood until completely dry. Cover the plates, wrap in foil, label with date and store at −20 °C for up to 4 weeks.& }1 o( _! h" T, `0 e
Laminin-coated six-well plate—Dilute laminin with fresh neural differentiation medium to a final concentration of 20 μg ml−1. Add 300 μl of laminin solution to each well of a sixwell plate. Allow the medium to hold as a big drop and spread within the central area of the well. Do not let the medium drain to the edge. Incubate the plate at 37 μC for 1 h. Laminin easily absorbs to plastic and tends to form aggregates at room temperature. Store laminin at −80 °C and thaw at 4 °C before using. Avoid aliquotting laminin into plastic tubes from the original glass vial.
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lhmxm

回复 千细胞 的帖子: K: @( X- c: A' h2 P. L

  {: h% |* u# i  x3 I7 W. y谢谢，非常好