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标题: 想用贴壁的方法培养神经干细胞?怎样包被圆盖片? [打印本页]

作者: lhmxm    时间: 2011-2-25 10:29     标题: 想用贴壁的方法培养神经干细胞?怎样包被圆盖片?

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?
作者: 千细胞    时间: 2011-2-25 15:36

可以考虑, e9 P* `+ \# Y; w  D9 ?

* R4 [2 l8 ]3 W0 E& m& M2 zEQUIPMENT SETUP
/ k4 K0 q8 u' X; j6 N; ^+ t0 Q! n. vPoly-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.
9 M% v0 r8 L: h# E  N1 B, B0 o, }2 }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.
: H& B9 ]6 _1 ?( i' s( F/ i/ T
作者: 千细胞    时间: 2011-2-25 15:36

可以考虑  @& C7 i3 T/ `6 n0 ?0 N. F
# v/ n( ^9 j$ {9 L0 ^; R9 K
EQUIPMENT SETUP
: D/ \3 \6 {% _" uPoly-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.
9 ]) I* J4 p7 n6 pLaminin-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.% `2 L7 k" F" y( x# Q

作者: lhmxm    时间: 2011-2-27 09:19

回复 千细胞 的帖子, O' i5 O: q# G4 Y" E' {8 S# w+ o1 S

; b7 o0 V4 V$ [谢谢,非常好




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