r******k
发帖数: 446 | 1 最近小弟要culture K562. 以前没杨过悬浮的细胞。大神们给点意见？
1. 复苏以后怎么又贴壁细胞？？？
2. 怎么传代呀。。。离心下来数细胞数？ 那传到25cm dish 和T75的dish都需要seed
多少细胞？ 多久算养好？
谢谢 |
A******y
发帖数: 2041 | 2 1. It will not attach to the plate. If you want the cells to attach to the
plate for immunostaining, you have to using fibronectin or matrigel etc.
2. The split is cells/mL. K562 should be sub-cultured at 200k/mL and no
more than 1 million/mL. However, it is very hard to kill K562 cells, so
even 1.5 to 2 million cells are possible. |
r******k
发帖数: 446 | 3 how often you change the medium?
the
【在 A******y 的大作中提到】
: 1. It will not attach to the plate. If you want the cells to attach to the
: plate for immunostaining, you have to using fibronectin or matrigel etc.
: 2. The split is cells/mL. K562 should be sub-cultured at 200k/mL and no
: more than 1 million/mL. However, it is very hard to kill K562 cells, so
: even 1.5 to 2 million cells are possible.
|
r******k
发帖数: 446 | 4 how often you change the medium?
the
【在 A******y 的大作中提到】
: 1. It will not attach to the plate. If you want the cells to attach to the
: plate for immunostaining, you have to using fibronectin or matrigel etc.
: 2. The split is cells/mL. K562 should be sub-cultured at 200k/mL and no
: more than 1 million/mL. However, it is very hard to kill K562 cells, so
: even 1.5 to 2 million cells are possible.
|
A******y
发帖数: 2041 | 5 2 to 3 days, it is better to count the cells though.
【在 r******k 的大作中提到】
: how often you change the medium?
:
: the
|
r******k
发帖数: 446 | 6 Thaaaaanks!
【在 A******y 的大作中提到】
: 2 to 3 days, it is better to count the cells though.
|
