产品名称：小鼠胚胎成纤维细胞-丝裂霉素处理

收到小鼠胚胎成纤维细胞-丝裂霉素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-丝裂霉素处理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 from ScienCell Research Laboratories are isolated from embryonic day 13 CD1 mouse embryos. These cells are cryopreserved at P0 and delivered frozen. Each vial contains 5 x 10^6 cells in 1 ml volume. MEF are characterized by immunofluorescence with antibody specific to fibronectin. MEF are negative for mycoplasma, bacteria, yeast, and fungi. MEF are guaranteed to further expand for 5 population doublings under the conditions provided by ScienCell Research Laboratories.

Recommended Medium

It is recommended to use DMEM (Cat. #09221) supplemented with 10% fetal bovine serum (FBS, Cat. #0010, 0025, 0500) for culturing MEF in vitro.

Product Use

MEF are used as a 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 Teratocarcinomas and Embryonic Stem Cells: A Practical Approach, (ed. E. J. Robertson), pp. 113-151. Oxford: IRL Press.

[2] Nagy et al. (2006) Preparing Mouse Embryo Fibroblasts Cold Spring Harbor Protocols. 2006: pdb.prot 4398.

小鼠胚胎成纤维细胞-丝裂霉素处理传代方法：
 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-丝裂霉素处理倒转放于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-丝裂霉素处理其它细胞系展示表：CHO-K1 中国仓鼠卵巢细胞 品牌ATCC 规格25毫升/株. 货期1-2周
COS-1 非洲绿肾 品牌ATCC 规格25毫升/株. 货期1-2周
COS-7 非洲绿猴肾细胞 品牌ATCC 规格25毫升/株. 货期1-2周
CV-1 猴肾细胞 品牌ATCC 规格25毫升/株. 货期1-2周
IAR20 小鼠肝细胞 品牌ATCC 规格25毫升/株. 货期1-2周
IEC-6 大鼠小肠隐窝上皮细胞 品牌ATCC 规格25毫升/株. 货期1-2周
LL-PK1, 猪肾细胞 品牌ATCC 规格25毫升/株. 货期1-2周
MC3T3-E1 小鼠胚胎成骨细胞 品牌ATCC 规格25毫升/株. 货期1-2周
MDCK 狗肾细胞 品牌ATCC 规格25毫升/株. 货期1-2周
MEF 小鼠胚胎成纤维细胞 品牌ATCC 规格25毫升/株. 货期1-2周
NIH3T3 小鼠胚胎成纤维细胞 品牌ATCC 规格25毫升/株. 货期1-2周
P815 小鼠肥大细胞 品牌ATCC 规格25毫升/株. 货期1-2周
PA12 小鼠成纤维细胞 品牌ATCC 规格25毫升/株. 货期1-2周
RTE 大鼠气管上皮细胞 品牌ATCC 规格25毫升/株. 货期1-2周
SF9 昆虫卵巢细胞 品牌ATCC 规格25毫升/株. 货期1-2周
SMC 兔主动脉平滑肌细胞 品牌ATCC 规格25毫升/株. 货期1-2周
Vero 猴肾细胞 品牌ATCC 规格25毫升/株. 货期1-2周
WEHI-3 小鼠血液细胞 品牌ATCC 规格25毫升/株. 货期1-2周
Y2 小鼠成纤维细胞 品牌ATCC 规格25毫升/株. 货期1-2周
L6 大鼠骨骼肌成肌细胞 品牌ATCC 规格25毫升/株. 货期1-2周
NRK 小鼠结缔组织细胞 品牌ATCC 规格25毫升/株. 货期1-2周
AtT-20 小鼠垂体瘤 品牌ATCC 规格25毫升/株. 货期1-2周
BC3H1, 鼠脑瘤 品牌ATCC 规格25毫升/株. 货期1-2周
BV-2 小鼠小胶质瘤细胞 品牌ATCC 规格25毫升/株. 货期1-2周
C6 大鼠脑胶质瘤 品牌ATCC 规格25毫升/株. 货期1-2周
Cyc-tAg 小鼠T**细胞瘤（SV40T抗原转染） 品牌ATCC 规格25毫升/株. 货期1-2周
EL-4 鼠T**细胞瘤 品牌ATCC 规格25毫升/株. 货期1-2周
FOX-NY 小鼠**瘤细胞 品牌ATCC 规格25毫升/株. 货期1-2周
GH3 大鼠垂体瘤细胞 品牌ATCC 规格25毫升/株. 货期1-2周