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细胞的复苏和传代
点击次数：159 发布时间：2016/12/21 15:44:31
细胞复苏:
1.应遵守慢冻快融的原则.先将水温锅调至37-37.5度,取出冻存的细胞迅速放入后交细胞面浸至水面以下不断摇动至融化.
2.在无菌台内将完全培养基加入50ml的小培养瓶内,约5ml左右,然后用无菌吸管从冻存管内取出细胞,置培养并内轻轻摇晃,使细胞均匀后置培养箱内培养.
细胞传代:
1.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.50ml培养瓶加入消化液约0.6ml,按此比例进行消化,(根据配制强度经验),晃动使消化液铺均匀置37度培养箱约2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.
2.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500rpm,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.
yWXD150 Homo sapiens, human
DSM 300 Sarcina ventriculi Goodsir
N-6; T 52A Nitrobacter sp.
yWXD4746 Homo sapiens, human
FGSC 1538 Neurospora crassa Shear et Dodge
TRTC 48882 [CBS 508.84, IMI 287300] Chaetomium robustum Ames, teleomorph
C [ATCC 25312, CIP 104337, NCIB 12416] Escherichia coli (Migula) Castellani ａnd Ch ..
NRRL YB-3239 [CBS 5762, CCRC 20470, FMJ 7135 .. Barnettozyma californica (Lodder) Kurtzman ..
CAL/CSF1 [CBS 6832, NRRL Y-12936, NRRL Y-829 .. Sporidiobolus salmonicolor Fell et Tallman
XM101 Dictyostelium discoideum Raper
yWXD2461 Homo sapiens, human
Patulibacter americanus Reddy ａnd Garcia-Pi ..
FAS21 Saccharomyces cerevisiae Meyen ex E.C. Hans ..
RP24238D19 Mus musculus, mouse
YJL065C BY4743, homozygous diploid [31359] Saccharomyces cerevisiae Meyen ex E.C. Hans ..
dried
4AB-65 Varicosporium elodeae Kegel
110S Ardenscatena maritimus
YGL218W BY4742, mating type alpha [16884] Saccharomyces cerevisiae Meyen ex E.C. Hans ..
FRI 182 Poria rhizomorpha Bagchee, teleomorph
int-2i Mus musculus, mouse
YPL227C BY4743, homozygous diploid [31065] Saccharomyces cerevisiae Meyen ex E.C. Hans ..
1.应遵守慢冻快融的原则.先将水温锅调至37-37.5度,取出冻存的细胞迅速放入后交细胞面浸至水面以下不断摇动至融化.
2.在无菌台内将完全培养基加入50ml的小培养瓶内,约5ml左右,然后用无菌吸管从冻存管内取出细胞,置培养并内轻轻摇晃,使细胞均匀后置培养箱内培养.
细胞传代:
1.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.50ml培养瓶加入消化液约0.6ml,按此比例进行消化,(根据配制强度经验),晃动使消化液铺均匀置37度培养箱约2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.
2.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500rpm,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.
yWXD150 Homo sapiens, human
DSM 300 Sarcina ventriculi Goodsir
N-6; T 52A Nitrobacter sp.
yWXD4746 Homo sapiens, human
FGSC 1538 Neurospora crassa Shear et Dodge
TRTC 48882 [CBS 508.84, IMI 287300] Chaetomium robustum Ames, teleomorph
C [ATCC 25312, CIP 104337, NCIB 12416] Escherichia coli (Migula) Castellani ａnd Ch ..
NRRL YB-3239 [CBS 5762, CCRC 20470, FMJ 7135 .. Barnettozyma californica (Lodder) Kurtzman ..
CAL/CSF1 [CBS 6832, NRRL Y-12936, NRRL Y-829 .. Sporidiobolus salmonicolor Fell et Tallman
XM101 Dictyostelium discoideum Raper
yWXD2461 Homo sapiens, human
Patulibacter americanus Reddy ａnd Garcia-Pi ..
FAS21 Saccharomyces cerevisiae Meyen ex E.C. Hans ..
RP24238D19 Mus musculus, mouse
YJL065C BY4743, homozygous diploid [31359] Saccharomyces cerevisiae Meyen ex E.C. Hans ..
dried
4AB-65 Varicosporium elodeae Kegel
110S Ardenscatena maritimus
YGL218W BY4742, mating type alpha [16884] Saccharomyces cerevisiae Meyen ex E.C. Hans ..
FRI 182 Poria rhizomorpha Bagchee, teleomorph
int-2i Mus musculus, mouse
YPL227C BY4743, homozygous diploid [31065] Saccharomyces cerevisiae Meyen ex E.C. Hans ..
