靶点科技-ROKEPIE细胞冬眠冷藏保存液

使用说明书

Product application/ intended use
ROKEPIE-FD01 is a supplement for your standard, well defined cell culture medium, enabling the hypothermic storage of cultured mammalian cells for several days. The lyophilized formulation should be dissolved in 2.0 mL solvent prior to use. This 2 mL of ROKEPIE-FD01 stock solution can be used to make 50 to 200 mL of hypothermic storage cell culture medium, depending on cell density and cell line in question.
About ROKEPIE-FD01
Research in the field of hibernation and its protective mechanisms led to the development of several novel chemical entities, designed to activate dormant protective mechanisms against the damage induced by hypothermia and subsequent rewarming. This product is designed to be used in your personal, well defined and cellspecific cell culture medium. Whereas other commercially available storage solutions completely replace cell culture 
medium, ROKEPIE-FD01 is simply an addition. Another benefit is that your cell culture medium immediately supports growth of viable cells after cold storage, without the need to remove the storage solution and apply fresh cell culture medium.
Storage and shipping
ROKEPIE-FD01 can be shipped and stored at room temperature. The container in which ROKEPIE-FD01 is delivered is meant to protect this product from light. Protect this product from direct light after reconstitution of the powder to prevent degradation of active substances. ROKEPIE-FD01 is delivered in a lyophilized formulation sufficient for 2.0 mL ROKEPIE-FD01 stock solution. The ROKEPIE-FD01 stock solution should be stored at 2 - 8 °C.
Directions for use
The following protocol is an example for use in adherent mammalian cells. The cell culture medium in this example uses bicarbonate as a pH-buffer. Cells are cultured in a flask with a standard phenolic style screw cap (i.e. without a membrane) which can be closed air-tight*. This enables 5% CO2 enrichedair to remain present in the flask during storage, preventing a sharp increase in pH due to a drop in CO2.
1. Clean the outside of the container with your disinfectant of choice. 
2. Dissolve the lyophilized ROKEPIE-FD01 in 2.0 mL solvent or cell culture medium of your choice. 
3. Add the ROKEPIE-FD01 stock solution in a 1:25 ratio (confluent monolayer) or 1:50 ratio (semi con-fluent monolayer) to either fresh- or already present cell culture medium.
4. Incubate the cells in the supplemented medium for a period of = 2 hours at 37°C .
5. Close the flask air-tight and place the cells in a cold-roomor refrigerator (4°C). 
6. After cold-storage, place cells in a humidified incubator with a slightly opened cap and replace the supplemented cell culture medium once cells have re-attached**. This normally takes about 6 – 24 hours. If cells have not detached during cold-storage, the supplemented cell culture medium can be replaced immediately. 
You can consult the schematic representation of the directions for use on the backside of this folder.
*) If no standard phenolic style caps are available, the filter can be covered by parafilm in order to seal the flask air-tight. Wells or dishes can be covered entirely with parafilm, or all around the edges of the lid. 
**) Detachment of cells can occur in some cell lines, but does not necessarily indicate they are apoptotic/ necrotic. ROKEPIEFD01 is not toxic at working concentrations and 24 hour exposure to cells at 37°C should not results in loss of viability. 
Warning and side notes
This product is for research use only. Even though this product has been tested on several cell lines (SMAC, HEK293, Skov-3, CaCo2), protection against the damage induced by hypothermia and rewarming by ROKEPIE-FD01 can be cell specific, and should therefore be tested on your cell-line of choice. It is recommended that the optimal duration of cold storage is determined per cell-line by the end-user. 
Contact and questions
Please tell us about your experiences with this product. Any information about protection by ROKEPIE-FD01 is appreciatedsince this product is still in a research stage of development. 
Do you need more information? Please visit our website for our FAQ or contact us through:


产品应用/预期用途
ROKEPIE-FD01可添加于标准培养基，如DMEM，1640培养基等，也可以用于个性化定制的培养基，从而可低温保存培养哺乳动物细胞数天。 冻干的形式提供，使用前，应将制剂溶于2.0 mL溶剂中。此2 mL ROKEPIE-FD01储备溶液可用于制备50至200 mL的低温储存细胞培养基，取决于所讨论的细胞密度和细胞系。
关于ROKEPIE-FD01
冬眠及其保护领域的研究机制导致了几种新型化学物质的发现，旨在激活休眠防止由以下因素引起的损害的保护机制体温过低和随后的变暖。这个产品是设计用于您的个性化，或定义明确且特定于细胞的细胞培养基。而其他商业可用的存储解决方案*替代了细胞培养介质，ROKEPIE-FD01只是添加。另一个好处是您的细胞培养基立即支持生长冷藏后的活细胞数量，无需移除储存溶液并应用新鲜的细胞培养基。
储存和运输
ROKEPIE-FD01可在室内运输和存放温度。运送ROKEPIE-FD01的容器旨在保护该产品免受光照。保护此产品配制后应避免直接光照活性物质的降解。 ROKEPIE-FD01交付于足以用于2.0 mL ROKEPIE-FD01的冻干制剂
储备溶液。 ROKEPIE-FD01库存溶液应为储存在2-8°C。
使用指南
以下协议是在粘附程序中使用的示例哺乳动物细胞。本例中的细胞培养基使用碳酸氢盐作为pH缓冲剂。将细胞在带盖的烧瓶中培养标准酚醛螺纹螺帽（即无膜）可以气密关闭*。这可以使5％的CO2富集储存期间空气会残留在烧瓶中，从而防止由于二氧化碳含量下降，pH值急剧上升。
1.用消毒剂清洁容器的外部，比如用75%的酒精。
2.将冻干的ROKEPIE-FD01溶解在2.0 mL溶剂中或您选择的细胞培养基。 
3.以1:25的比例添加ROKEPIE-FD01储备溶液（汇合单层）或1:50比率（半汇合单层）到新鲜或已经存在的细胞培养中。
4.将细胞在补充培养基中孵育在37°C下大约 2小时。
5.密闭培养瓶，将细胞放在冷室中或冰箱（摄氏4度）。
6.冷藏后，将细胞放在潮湿的培养箱中用稍微打开的帽子，并替换补充细胞培养基一旦细胞重新附着。**这个通常大约需要6到24小时。如果细胞没有在冷藏过程中分离，补充的细胞培养基可以立即更换。
您可以查阅说明的示意图在此文件夹的背面使用。
*）如果没有标准的酚醛塑料瓶盖，则过滤器可以用封口膜覆盖，以将培养瓶密封。孔或皿可以*用封口膜覆盖，或全部盖的边缘周围。
**）在某些细胞系中可能发生细胞分离，但确实如此不一定表示它们是凋亡/坏死的。 ROKEPIE FD01在工作浓度和24小时无毒暴露于37°C的细胞不应导致丧失生存能力。
警告和注意事项
本产品仅供研究使用。即使这个ROKEPIE-FD01产品已在多种细胞系（SMAC，HEK293，Skov-3，CaCo2）使用来防止体温过低和回暖可能导致的细胞伤害，但是由于每个细胞的特性，因此在您的选择时。建议optimal的冷藏持续时间存储是由终用户按每个细胞预实验来确定的。
问题咨询
请告诉我们您使用此产品的经验。任何有关ROKEPIE-FD01使用的信息，我们都很重视，因为该产品仍处于进一步研发优化阶段。你需要更多的信息？请访问我们的网站。