靶點科技-ROKEPIE細胞冬眠冷藏保存液
使用說明書
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:
產品應用/預期用途
ROKEPIE-FD01可添加于標準培養基，如DMEM，1640培養基等，也可以用于個性化定制的培養基，從而可低溫保存培養哺乳動物細胞數天。 凍干的形式提供，使用前，應將制劑溶于2.0 mL溶劑中。此2 mL ROKEPIE-FD01儲備溶液可用于制備50至200 mL的低溫儲存細胞培養基，取決于所討論的細胞密度和細胞系。
關于ROKEPIE-FD01
冬眠及其保護領域的研究機制導致了幾種新型化學物質的發現，旨在激活休眠防止由以下因素引起的損害的保護機制體溫過低和隨后的變暖。這個產品是設計用于您的個性化，或定義明確且特定于細胞的細胞培養基。而其他商業可用的存儲解決方案*替代了細胞培養介質，ROKEPIE-FD01只是添加。另一個好處是您的細胞培養基立即支持生長冷藏后的活細胞數量，無需移除儲存溶液并應用新鮮的細胞培養基。
儲存和運輸
ROKEPIE-FD01可在室內運輸和存放溫度。運送ROKEPIE-FD01的容器旨在保護該產品免受光照。保護此產品配制后應避免直接光照活性物質的降解。 ROKEPIE-FD01交付于足以用于2.0 mL ROKEPIE-FD01的凍干制劑
儲備溶液。 ROKEPIE-FD01庫存溶液應為儲存在2-8°C。
使用指南
以下協議是在粘附程序中使用的示例哺乳動物細胞。本例中的細胞培養基使用碳酸氫鹽作為pH緩沖劑。將細胞在帶蓋的燒瓶中培養標準酚醛螺紋螺帽（即無膜）可以氣密關閉*。這可以使5％的CO2富集儲存期間空氣會殘留在燒瓶中，從而防止由于二氧化碳含量下降，pH值急劇上升。
1.用消毒劑清潔容器的外部，比如用75%的酒精。
2.將凍干的ROKEPIE-FD01溶解在2.0 mL溶劑中或您選擇的細胞培養基。
3.以1:25的比例添加ROKEPIE-FD01儲備溶液（匯合單層）或1:50比率（半匯合單層）到新鮮或已經存在的細胞培養中。
4.將細胞在補充培養基中孵育在37°C下大約 2小時。
5.密閉培養瓶，將細胞放在冷室中或冰箱（攝氏4度）。
6.冷藏后，將細胞放在潮濕的培養箱中用稍微打開的帽子，并替換補充細胞培養基一旦細胞重新附著。**這個通常大約需要6到24小時。如果細胞沒有在冷藏過程中分離，補充的細胞培養基可以立即更換。
您可以查閱說明的示意圖在此文件夾的背面使用。
*）如果沒有標準的酚醛塑料瓶蓋，則過濾器可以用封口膜覆蓋，以將培養瓶密封。孔或皿可以*用封口膜覆蓋，或全部蓋的邊緣周圍。
**）在某些細胞系中可能發生細胞分離，但確實如此不一定表示它們是凋亡/壞死的。 ROKEPIE FD01在工作濃度和24小時無毒暴露于37°C的細胞不應導致喪失生存能力。
警告和注意事項
本產品僅供研究使用。即使這個ROKEPIE-FD01產品已在多種細胞系（SMAC，HEK293，Skov-3，CaCo2）使用來防止體溫過低和回暖可能導致的細胞傷害，但是由于每個細胞的特性，因此在您的選擇時。建議optimal的冷藏持續時間存儲是由終用戶按每個細胞預實驗來確定的。
問題咨詢
請告訴我們您使用此產品的經驗。任何有關ROKEPIE-FD01使用的信息，我們都很重視，因為該產品仍處于進一步研發優化階段。你需要更多的信息？請訪問我們的網站。
