产品名称：小鼠胚胎成纤维细胞--CF1小鼠丝裂霉素C处理

收到小鼠胚胎成纤维细胞--CF1小鼠丝裂霉素C处理时如无异常情况，请在显微镜下观察细胞密度，如为贴壁细胞，未超过80%汇合度时，将培养瓶中培养液吸出，留下10ml培养液继续培养；超过80%汇合度时，请按细胞培养条件传代培养。如为悬浮细胞，吸出培养液、1000转/分钟离心2分钟，吸出上清，管底细胞用新鲜培养基悬浮细胞后移回培养瓶。

小鼠胚胎成纤维细胞--CF1小鼠丝裂霉素C处理Description

Mouse Embryonic Fibroblasts (MEF) are used to support the growth of mouse and human pluripotent stem cells [1]. MEF not only provide a substrate for pluripotent stem cells to grow on, but also secrete critical growth factors to maintain stem cell pluripotency. MEF are isolated from mouse embryos and used at early passages [2]. To serve as feeder cells, MEF must be treated with mitomycin C or by irradiation to prevent cell proliferation. The treated cells can also be used to generate conditioned medium for feeder-free culture of pluripotent stem cells

MEF-mt from ScienCell Research Laboratories are isolated from embryonic day 13 CF1 mouse embryos. These cells have been treated with mitomycin C to prevent further cell division. They are cryopreserved at passage 3 and delivered frozen. Each vial contains 1 x 10^6 cells in 1 ml volume. MEF-mt are characterized by immunofluorescence with antibody specific to fibronectin. MEF-mt are negative for mycoplasma, bacteria, yeast, and fungi. MEF are guaranteed to further culture under the conditions provided by ScienCell Research Laboratories; however, MEF-mt are not recommended for expanding as mitomycin C treatment prevents further cell proliferation.

Recommended Medium

It is recommended to use DMEM (Cat. No. 09221) containing 10% fetal bovine serum (FBS, Cat. No. 0010, 0025, 0500) for the culturing of MEF-mt cells in vitro.

Product Use

MEF-mt are used as feeder layer in mouse and human pluripotent stem cell culture. They are for research use only. They are not approved for human or animal use, or for application in in vitro diagnostic procedures.

Storage

Upon receiving, directly and immediately transfer the cells from dry ice to liquid nitrogen and keep the cells in liquid nitrogen until they are needed for experiments.

Shipping

Dry ice.

Reference

[1] Bradley A. (1987) “Production and analysis of chimaeras.” In Robertson EJ, Teratocarcinomas and Embryonic Stem Cells: A Practical Approach (pp 113-51). Oxford: IRL Press.

[2] Nagy A, Gertsenstein M, Vintersten K, Behringer R. (2006) “Preparing Mouse Embryo Fibroblasts.” Cold Spring Harbor Protocols. pdb.prot 4398.

小鼠胚胎成纤维细胞--CF1小鼠丝裂霉素C处理传代方法：
 收到细胞后，取出培养瓶在倒置显微镜下观察细胞生长情况。
（一）如果细胞未长满，用75%酒精喷洒整个瓶**后放到超菌台内，严格无菌操作，打开细胞培养瓶，吸出培养液，仅留下10ml培养液在瓶内继续培养。
(二)如果细胞已长满，即可进行传代培养。具体步骤如下：
1. 弃去培养液，用PBS（不含钙，镁离子）洗1-2次。
2. 加1ml消化液（0.25%Trypsin-0.53mM EDTA）于培养瓶中，小鼠胚胎成纤维细胞--CF1小鼠丝裂霉素C处理倒转放于37度培养箱1-3分钟预热，然后又将培养瓶倒转大约30秒后，在倒置显微镜下观察细胞消化情况，若细胞大部分变圆分散，轻敲几下培养培养瓶，细胞随即脱落下来。
3. 加入6-8ml完全培养基，吸出，分到新的培养瓶中。1:3~1:6传代；2~3天1次。 
。
注意：传代后一半用我们的培养基，一半用你们的，以免细胞不适应而造
成生长不好。
冻存方法：   冻存液：基础培养基+5%DMSO+20%FBS 
储存：液氮储存
小鼠胚胎成纤维细胞--CF1小鼠丝裂霉素C处理其它细胞系展示表：yb-axbz-202 C3H小鼠骨肉瘤 LM8 规格25毫升/株.
yb-axbz-203 大鼠骨肉瘤 VMR106 规格25毫升/株.
yb-axbz-204 猴肾C M145 规格25毫升/株.
yb-axbz-205 兔成骨C IF 规格25毫升/株.
编号 中文 英文名称 规格25毫升/株.
yb-axbz-206 小鼠白血病 L1210 规格25毫升/株.
yb-axbz-207 大鼠皮层神经元细胞 RN-c 规格25毫升/株.
yb-axbz-208 大鼠主动脉平滑肌细胞 RSMC 规格25毫升/株.
yb-axbz-209 小鼠小胶质瘤细胞 Bv-2 规格25毫升/株.
yb-axbz-210 鼠杂交骨髓瘤细胞 K6H6/B5 规格25毫升/株.
yb-axbz-211 大鼠脑膜细胞 RMC 规格25毫升/株.
yb-axbz-212 转基因细胞 3T3/Fas/com 规格25毫升/株.
yb-axbz-213 大鼠肝癌细胞 CRL-1548 规格25毫升/株.
yb-axbz-214 小鼠T**细胞瘤细胞 Cyc-Tag(S49) 规格25毫升/株.
yb-axbz-215 小鼠骨髓细胞 FDC-P1 规格25毫升/株.
yb-axbz-216 小鼠垂体瘤细胞 GT1-1 规格25毫升/株.
yb-axbz-217 小鼠神经母细胞瘤细胞与大鼠胶质瘤细胞之融合细胞 NG108-15 规格25毫升/株.
yb-axbz-218 小鼠胚胎成纤维细胞(来自NIH3T3) PA12 规格25毫升/株.
yb-axbz-219 小鼠血细胞 WEHI-3 规格25毫升/株.
yb-axbz-220 小鼠红白血病细胞 MEL 规格25毫升/株.
yb-axbz-221 兔主动脉平滑肌细胞 SMC 规格25毫升/株.
yb-axbz-222 小鼠垂体瘤细胞(分泌促生长**分泌**) AtT-20 规格25毫升/株.
yb-axbz-223 小鼠骨髓瘤细胞 Fox-NY 规格25毫升/株.
yb-axbz-224 兔主动脉平滑肌细胞 CCC-SMC-1 规格25毫升/株.
yb-axbz-225 兔主动脉平滑肌细胞 CCC-SMC-2 规格25毫升/株.
yb-axbz-226 大鼠垂体瘤细胞 MMQ 规格25毫升/株.
yb-axbz-227 小鼠原B细胞株 BaF3 规格25毫升/株.
yb-axbz-228 小鼠脑瘤细胞 BC3H1 规格25毫升/株.