【正常肝细胞】L-02细胞【正常肝细胞】L-02细胞贴壁细胞: 对于贴壁细胞应先吸(倒)尽培养基，吸的越干净越好，以免中和后加入的消化液，使强度减弱（或PBS洗1－3次）。50ml培养瓶加入消化液约1-3ml，按此比例进行消化，(根据经验)，晃动使消化液铺均匀置37度培养箱约2-5分钟，镜下见细胞收缩变圆或少数脱落后，轻轻振动瓶底使细胞全部脱落，加入2-3ml完全培养基后，轻轻吹打，使细胞基本成单个悬浮，然后分置其它无菌培养瓶内，加入完全培养基后继续培养或实验。【正常肝细胞】L-02细胞培【正常肝细胞】L-02细胞培悬浮细胞: 一般传代可直接将细胞原液分置其它培养瓶内，加入完全培养基继续培养，如要高浓度可先离心1000rpm，5min后加入完全培养基，轻轻吹匀后，分置其它培养瓶内加入完全培养基继续培养。需要指出的是，在不同的科研小组中这些术语的定义会有所不同。许多研究员不用细胞系这个词表示细胞群除非细胞经过了遗传改造。
【正常肝细胞】L-02细胞培养基的配制【正常肝细胞】L-02细胞培养基的配制

培养基	品牌	货号
D-MEM/F-12	GIBCO	12400024
Dulbecco's Modified Eagle Medium (D-MEM), powder (high glucose)	GIBCO	12800017
F-12 Nutrient Mixture (Ham) powder	GIBCO	21700075
Iscove's Modified Dulbecco's Medium (IMDM) powder	GIBCO	12200036
Leibovitz's L-15 Medium powder	GIBCO	41300039
McCOY's 5A	SIGMA	M4892
Minimum Essential Medium (MEM) Alpha Medium powder (MEMα) with ribonucleosides and deoxyribonucleosides	GIBCO	11900024
Minimum Essential Medium (MEM) Alpha Medium powder (MEMα) without ribonucleosides and deoxyribonucleosides	GIBCO	12000022
Minimum Essential Medium (MEM) powder	GIBCO	41500034
NUTRIENT MIXTURE F12 HAM KAIGHN'S MODIFICATION (F12K)	SIGMA	N3520
RPMI Medium 1640	GIBCO	31800022
EGF	SIGMA	E9644
Sodium pyruvate	SIGMA	P2256-25G
MEDIUM 199, WITH EARLE'S SALTS AND L-GLUTAMINE, WITHOUT SODIUM BICARBONATE. CELL CULTURE TESTED	SIGMA	M5017

培养基品牌货号D-MEM/F-12GIBCO12400024Dulbecco's Modified Eagle Medium (D-MEM), powder (high glucose)GIBCO12800017F-12 Nutrient Mixture (Ham) powderGIBCO21700075Iscove's Modified Dulbecco's Medium (IMDM) powderGIBCO12200036Leibovitz's L-15 Medium powderGIBCO41300039McCOY's 5A

SIGMA

M4892Minimum Essential Medium (MEM) Alpha Medium powder (MEMα)
with ribonucleosides and deoxyribonucleosidesGIBCO11900024Minimum Essential Medium (MEM) Alpha Medium powder (MEMα)
without ribonucleosides and deoxyribonucleosidesGIBCO12000022Minimum Essential Medium (MEM) powderGIBCO41500034NUTRIENT MIXTURE F12 HAM KAIGHN'S
MODIFICATION (F12K)SIGMAN3520RPMI Medium 1640GIBCO31800022EGFSIGMAE9644Sodium pyruvateSIGMAP2256-25GMEDIUM 199, WITH EARLE'S SALTS AND L-GLUTAMINE, WITHOUT SODIUM BICARBONATE. CELL CULTURE TESTEDSIGMAM5017培养基品牌货号培养基培养基品牌品牌货号货号D-MEM/F-12GIBCO12400024D-MEM/F-12D-MEM/F-12GIBCOGIBCO1240002412400024Dulbecco's Modified Eagle Medium (D-MEM), powder (high glucose)GIBCO12800017Dulbecco's Modified Eagle Medium (D-MEM), powder (high glucose)Dulbecco's Modified Eagle Medium (D-MEM), powder (high glucose)GIBCOGIBCO1280001712800017F-12 Nutrient Mixture (Ham) powderGIBCO21700075F-12 Nutrient Mixture (Ham) powderF-12 Nutrient Mixture (Ham) powderGIBCOGIBCO2170007521700075Iscove's Modified Dulbecco's Medium (IMDM) powderGIBCO12200036Iscove's Modified Dulbecco's Medium (IMDM) powderIscove's Modified Dulbecco's Medium (IMDM) powderGIBCOGIBCO1220003612200036Leibovitz's L-15 Medium powderGIBCO41300039Leibovitz's L-15 Medium powderLeibovitz's L-15 Medium powderGIBCOGIBCO4130003941300039McCOY's 5A

SIGMA

M4892McCOY's 5AMcCOY's 5A

SIGMA

SIGMA

SIGMASIGMASIGMASIGMAM4892M4892Minimum Essential Medium (MEM) Alpha Medium powder (MEMα)
with ribonucleosides and deoxyribonucleosidesGIBCO11900024Minimum Essential Medium (MEM) Alpha Medium powder (MEMα)
with ribonucleosides and deoxyribonucleosidesMinimum Essential Medium (MEM) Alpha Medium powder (MEMα)
with ribonucleosides and deoxyribonucleosidesGIBCOGIBCO1190002411900024Minimum Essential Medium (MEM) Alpha Medium powder (MEMα)
without ribonucleosides and deoxyribonucleosidesGIBCO12000022Minimum Essential Medium (MEM) Alpha Medium powder (MEMα)
without ribonucleosides and deoxyribonucleosidesMinimum Essential Medium (MEM) Alpha Medium powder (MEMα)
without ribonucleosides and deoxyribonucleosidesGIBCOGIBCO1200002212000022Minimum Essential Medium (MEM) powderGIBCO41500034Minimum Essential Medium (MEM) powderMinimum Essential Medium (MEM) powderGIBCOGIBCO4150003441500034NUTRIENT MIXTURE F12 HAM KAIGHN'S
MODIFICATION (F12K)SIGMAN3520NUTRIENT MIXTURE F12 HAM KAIGHN'S
MODIFICATION (F12K)NUTRIENT MIXTURE F12 HAM KAIGHN'S
MODIFICATION (F12K)SIGMASIGMAN3520N3520RPMI Medium 1640GIBCO31800022RPMI Medium 1640RPMI Medium 1640GIBCOGIBCO3180002231800022EGFSIGMAE9644EGFEGFSIGMASIGMAE9644E9644Sodium pyruvateSIGMAP2256-25GSodium pyruvateSodium pyruvateSIGMASIGMAP2256-25GP2256-25GMEDIUM 199, WITH EARLE'S SALTS AND L-GLUTAMINE, WITHOUT SODIUM BICARBONATE. CELL CULTURE TESTEDSIGMAM5017MEDIUM 199, WITH EARLE'S SALTS AND L-GLUTAMINE, WITHOUT SODIUM BICARBONATE. CELL CULTURE TESTEDMEDIUM 199, WITH EARLE'S SALTS AND L-GLUTAMINE, WITHOUT SODIUM BICARBONATE. CELL CULTURE TESTEDSIGMASIGMAM5017M5017 2、 按要求添加碳酸氢钠、谷氨酸钠、HEPES等，充分搅拌使之溶解。
3、 调pH至7.2左右。
4、 加水至最终体积。
5、 在超净台中对溶液进行滤过除菌，分装入250 mL或500 mL玻璃瓶中，用翻帽塞塞紧瓶口。
6、 瓶口封好，4℃冰箱贮存。
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