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pc12细胞/菌种
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价格：￥电议
更新日期：2026/9/8 3:56:46
生 产 地：美洲
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详细内容
1，pc12细胞的复苏和培养
从液氮罐中取出盛有pc12细胞的塑料螺口小瓶，立即投入37℃温水中，并不时摇动令其尽快融化，然后从37℃水浴中取出塑料螺口小瓶，用酒精消毒后用吸管吸出细胞悬浮液，注入离心管并加入10倍培养液，混合后离心(一千转每分钟，五分钟)除去上清液，再重复用培养液洗一次。然后用培养液稀释成1x十的四次方个每毫升密度的细胞悬液，接种于75平方厘米塑料培养瓶中，每皿十毫升，置于37度含百分之十二氧化碳的培养箱内进行培养。培养液由百分之九十的DMEM、百分之五胎牛血清、百分之五马血清、0.01%谷氨酰胺组成。
2，pc12细胞的扩增
pc12细胞培养5~10天，经0.25胰蛋白酶消化(37度，五分钟)传代，以1x十的四次方个每毫升密度接种于75平方厘米塑料培养瓶中进行扩增培养。
3，体外诱导pc12细胞向交感神经元样细胞分化
取传五代以上pc12细胞，经消化分散后，制成5x十的五次方每毫升密度的细胞悬液，接种于涂有小牛皮胶的35mm塑料培养皿中，没皿2ml，置于36度，10%co2的培养箱内进行培养。培养液由90%DMEM，5%马血清、NGF20ng每毫升和谷氨酰胺0.10g/L组成。每周换液两次，每次更换一半新鲜培养液。
(二)结果
1，pc12细胞 的生长发育和形态学特征
pc12细胞种植后8小时，生长良好的pc12细胞开始分裂增殖并凝聚成团生长，pc12细胞呈圆形，胞体周围有晕光。培养5~10天后，分裂增殖的pc12细胞已形成密集的细胞簇，布满整个培养瓶。
2，体外诱导pc12细胞向交感神经元样细胞分化
分散的pc12细胞在含NGF的培养条件培养3天后，pc12细胞开始停止分裂。并逐渐分化为具有交感神经元特征的细胞，开始长出神经突起，培养5天后，大多数pc12细胞转变为类似于交感神经元的形态，突起逐渐增多并延长，形成稀疏的网络。随着培养时间的延长，交感神经元样细胞的胞体逐渐增大，胞体主干和分支逐渐延长并增粗，经NSE免疫组织化学染色呈阳性反应。
一、细胞信息
| Organism | Rattus norvegicus, rat |
|---|---|
| Tissue | adrenal gland |
| Product Format | frozen |
| Morphology | small irregularly shaped cells |
| Culture Properties | floating clusters; few scattered lightly attached cells. |
| Biosafety Level | 1 Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| Disease | pheochromocytoma |
| Gender | male |
| Applications | This cell line is a suitable transfection host. |
| Storage Conditions | liquid nitrogen vapor phase |
二、细胞特点
| Karyotype | 40 chromosomes; 38 autosomes plus XY Ref |
|---|---|
| Images |  |
| Derivation | The PC-12 cell line was derived from a transplantable rat pheochromocytoma. |
| Clinical Data | male |
| Receptor Expression | nerve growth factor (NGF), expressed |
| Genes Expressed | catecholamines; dopamine; norepinephrine |
| Cellular Products | catecholamines; dopamine; norepinephrine |
| Tumorigenic | Yes |
| Effects | Yes, in New England Deaconess Hospital strain rats |
| Comments | The cells respond reversibly to NGF by induction of the neuronal phenotype when plated on Collagen IV coated culture flasks. The cells do not synthesize epinephrine. |
三、培养方法
| Complete Growth Medium | The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, Catalog No. 30-2001. To make the complete growth medium, add the following components to the base medium:
|
|---|---|
| Subculturing | Protocol: Volumes used for this protocol are for a 75cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Corning® T⁻⁷5 flasks (catalog #431464) are recommended for subculturing this product.
Medium Renewal: Every 2 to 3 days |
| Cryopreservation | Freeze medium: Complete growth medium supplemented with 10% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase |
| Culture Conditions | Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C |
四、发展历史
| Population Doubling Time | 48 hrs |
|---|
