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evaleiyuhang / parkinson

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nas2 ipsc pd 上 #6

Open evaleiyuhang opened 11 months ago

evaleiyuhang commented 11 months ago

cell，PDO3，PDO6，3个6w，1个freeze，1个split到3个6w，1个split到3个6w和2个12w。 【Split1:3】：remove，+pbs-，+EDTA 3min RT-，准备3个6w+E8 1ml，+E8 1ml 3次 split 3个6w。 【Split 1:4】：remove，+pbs-，+EDTA 3min-，准备2个12w+E8和3个6w+E8，+E8 1ml 至2个6w，+E8 1ml至1个6w和2个12w。 【1个6w freeze】：PBs，EDTA，准备cry，+E8 1ml 1000rpm 3min,remove，+cry 500ul，freeze-80

evaleiyuhang commented 11 months ago

remove，+pbs-，+aktais 4min-，+n2b2f 2ml分给2个6w， new coat board，remove，+n2b2f 1ml，+rc 100ul，+cell1ml（split pod3（2个6w到4个6w），pod6（2个6w到4个6w））

evaleiyuhang commented 11 months ago

"remove，+pbs-，+3ml n2b2f（4个6w，4个6w） remove，+pbs-，+3ml e8（六孔板12个孔）"

evaleiyuhang commented 11 months ago

split1：3，PDO3有2个6w其中一个6w to 3个6w，PDO6有2个6w其中一个6w to 3个6w。

【split】coat 6个6w（remove，+n2b2f 1ml）cell（remove，+pbs-，+aktis 4min-，+n2b2f 1ml3次 分给3个6w）

evaleiyuhang commented 11 months ago

"Pd3，: Split 1to 4， 1to 6

  1. coat，+e8 2ml
  2. PBS、EDTA， 5min
  3. Split 1to 4， 1to 6，一半加EGF2，一半不加 pd6: Split 1to 4， 1to 6
  4. coat，+e8 2ml
  5. PBS、EDTA， 5min Split 1to 4， 1to 6，一半加EGF2，一半不加"
evaleiyuhang commented 11 months ago

"Pd3，: 转移？ pd6: ？ 17:30-17:40🥥

"

evaleiyuhang commented 11 months ago

"Feed cells 7x6w PD3; 7x6w,PD6 16:30-17:00🥥

Feed cells 7x6w PD3; 7x6w,PD6

  1. Remove medium 95%
  2. +5ml e8(50ml complete E8: 50ml E8+1ml E8 supplements) +5ul FGF2(1 in 1000)"
evaleiyuhang commented 11 months ago

" NAS2 ipsc PD8 上层，4X6well：2X6w split1:3，2X6w keep 16/11/2023 18:06 🥥 酒精喷所有放通风橱 【2X6w 1:3 6X6w（cell repalatintes）】： Remove all media in 2X6w one by one，wash with PBS 1ml，+EDTA 1ml 4min -， Add 2.4ml E8 to cell repalantintes 命名（NAS2 ipsc PD8 16/11/23） collect cells with E8 2ml，split to 3X6w cell repalantine Clean 2X6w cell plate with PBS 1ml 【2X6w feed】： Remove all media，+2ml E8 酒精消毒上层恒温箱，分开放"