AML12细胞系

参考资料：
https://www.atcc.org/products/all/CRL-2254.aspx#generalinformation

CRL-2254 Low High.jpg

Applications：

This cell line is a suitable transfection host.
该细胞系适宜转染。

Derivation：

The AML12 (alpha mouse liver 12) cell line was established from hepatocytes from a mouse (CD1 strain, line MT42) transgenic for human TGF alpha.
AML12(alpha小鼠肝12)细胞系来源于(CD1，MT42品系)人源TGF -alpha转基因小鼠的的肝实质细胞。

Genes Expressed

albumin; human transforming growth factor alpha (TGF alpha); mouse TGF alpha.
白蛋白;人源转化生长因子(TGF);鼠源转化生长因子α。

Cellular Products

albumin; human transforming growth factor alpha (TGF alpha); mouse TGF alpha.
白蛋白;人源转化生长因子(TGFα);鼠源TGFα。

Effects

No, the cells do not form colonies or grow in soft agar.
这些细胞不会形成克隆，也不会在软琼脂中生长。
No, the cells were not tumorigenic in immunosuppressed mice.
这些细胞在免疫抑制小鼠中不具有致瘤性。

Comments

The AML12 (alpha mouse liver 12) cell line was established from hepatocytes from a mouse (CD1 strain, line MT42) transgenic for human TGF alpha.
AML12(alpha小鼠肝12)细胞系来源于(CD1，MT42品系)人源TGF -alpha转基因小鼠的的肝实质细胞。
By electron microscopy, these cells exhibit typical hepatocyte features such as peroxisomes and bile canalicular like structure.
通过电镜观察，这些细胞表现出典型的肝细胞特征，如过氧化物酶体和胆管样结构。
AML12 cells retain the capacity to express high levels of mRNA for serum (albumin, alpha 1 antitrypsin and transferrin) and gap junction (connexins 26 and 32) proteins, and contain solely isoenzyme 5 of lactate dehydrogenase.
AML12细胞保留了高水平(血清白蛋白、α1抗胰蛋白酶和转铁蛋白)和缝隙连接(连接蛋白26和32)蛋白mRNA的能力，并且仅含有乳酸脱氢酶同工酶5。
The cells express high levels of human TGF alpha and lower levels of mouse TGF alpha.
细胞表达高水平的人TGFα，低水平的小鼠TGFα。
Expression of liver specific proteins decreases with time in culture, but is reactivated by growing the cells in serum free medium.
肝脏特异性蛋白的表达随培养时间的延长而降低，但在无血清培养基中通过细胞生长而重新激活。

Complete Growth Medium

The base medium for this cell line is DMEM:F12 Medium (ATCC 30-2006). To make the complete growth medium, add the following component to the 500 mL of the base medium:
该细胞系的基础培养基是DMEM：F12。要制作完全培养基，请将以下成分添加到500 mL的基础培养基中：

Supplemented with:
10% fetal bovine serum (FBS; ATCC 30-2020)
10 µg/ml insulin胰岛素
5.5 µg/ml transferrin转铁蛋白
5 ng/ml selenium硒
40 ng/ml dexamethasone地塞米松

This medium is formulated for use with a 5% CO2 in air atmosphere.
该培养基可在空气中通入5％CO 2使用。
Subculturing传代
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
该方案中使用的容积为75 cm2培养瓶；其他尺寸培养皿，按比例减少或增加培养基量。

1.Remove and discard culture medium.除去培养基
2.Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
用0.25％（w / v）胰蛋白酶-0.53 mM EDTA溶液短暂冲洗细胞层，以除去所有痕量的含胰蛋白酶抑制剂的血清。
3.Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
向烧瓶中加入2.0至3.0 ml的胰蛋白酶-EDTA溶液，并在倒置显微镜下观察细胞直至细胞层分散（通常在5至15分钟内）。
Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
注意：为避免结块，在消化细胞时，请勿通过敲击或摇动烧瓶来搅动细胞。难以消化的细胞可以放置在37°C培养箱以便于消化。
4.Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
加入6.0至8.0 ml完全生长培养基，并轻轻吸取细胞。
5.Add appropriate aliquots of the cell suspension to new culture vessels.
将细胞悬液的等分试样添加到新的培养皿中。
6.Incubate cultures at 37°C.
在37°C下继续培养。

Subcultivation Ratio: 1:4 to 1:6
传代比例：1:4 - 1:6
Medium Renewal: 2 to 3 times a week.
培养基更换频率：一周2 - 3次
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.

Cryopreservation细胞冻存

Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
冻存培养基：完全生长培养基，添加5％（v / v）DMSO
Storage temperature: liquid nitrogen vapor phase
储存温度：液氮

Culture Conditions培养条件

Temperature: 37°C