Transport Mode
Liquid Nitrogen Transportation
1、Adsorptive Liquid Nitrogen Transportation ,No free liquid nitrogen ,White vapor emission indicates normal operation.
2、Temperature monitoring devices should closely monitor the temperature inside the tank during transportation. If any abnormalities are detected, you can copy the PDF and Excel data from the temperature recorder. (One end of the temperature monitoring device can be unplugged to reveal a USB connector. Simply insert it into a computer to copy the data. If you encounter any difficulties, you can contact the sales staff and technical support for assistance.)
3、Alloy Combination Lock Designed to ensure no third-party access to the LN₂ tank or cell samples during transportation from dispatch to receipt, thereby safeguarding cell integrity.
4、GPS Tracker Enables real-time tracking of cell transportation routes to prevent loss.
5、Liquid Nitrogen Tank, temperature monitoring device, alloy combination lock, and GPS tracker are returnable components. Please store them properly and avoid damage; failure to comply will result in blacklisting.
Reference Citation
blush.
Scope of Application
Renal tubular epithelial cells are used to investigate the mechanisms of acute and chronic kidney diseases (such as acute kidney injury, chronic kidney disease), assess drug-induced nephrotoxicity, study the progression of renal fibrosis, explore mechanisms of cellular regeneration and repair, and examine metabolic disorders (e.g., diabetic nephropathy) and regulation of signaling pathways (such as TGF-β, Wnt). They also serve as disease models and drug screening platforms.
II 试剂与材料
Ø 冻存原代肾皮质细胞（Cat# LV-PRC001/2/3/4）
Ø 肾皮质细胞培养基（DMEM/F12+10%FBS+1%P/S）
Ø 胶原包被培养瓶（LV-coated）
Ø 无菌15ml离心管
Ø 一次性移液管
Ø 37℃/5%CO2培养箱
Ø 移液枪
Ø 恒温水浴锅（37℃预热）
Ø 水平离心机（带水平转子，可离心15ml离心管）
Ø 生物安全柜
Ø 宽口移液枪头（普通移液枪头剪去尖头，再灭菌使用）
Ø 75%酒精
Ø 0.25%胰酶
Ø PBS
重要提示：解冻时，冻存细胞转移必须使用液
氮转移；在整个复苏实验过程中必须全程使用
宽口枪头。
III 细胞的复苏与铺板
1.原代肾皮质细胞培养基放入37℃恒温水浴锅中充分预热，分出其中10ml加入到15ml离心管中并置于37℃水浴锅中充分预热15min。。
2.将冻存的细胞从冷藏位置经液氮迅速转移至37℃恒温水浴锅中。将冻存管尽可能多的浸入37℃水中，顺时针水平摇动，但必须确保冻存管管盖保持在水
面以上。
3.解冻冻存管约90~120s，至冻存管中只有少量碎冰漂浮即可。
4.用75%酒精消毒冻存管，并将其转移到生物安全柜。
5.用宽口枪头将肾皮质细胞重悬（轻轻吹打2次），并将细胞悬液逐滴加入含有10mL 预热的肾皮质细胞培养基的15ml离心管中（注意：冻存管与枪头上残留细胞，可用1ml肾皮质细胞培养基润洗冻存管与枪头）。
6.盖上盖子，将离心管轻微上下颠倒3次混匀。500g，常温离心5min，去上清并用培养基重悬。
7.沉淀的细胞用肾皮质细胞培养基重悬并定容至4ml，用台盼蓝排除法测定细胞的活力和细胞总量。
8.将细胞按2×10^4cells/cm2接种至胶原包被培养瓶中。摇匀，在37℃/5%CO2培养箱中培养，48h后换液。
IV 肾皮质细胞培养与传代
1.肾皮质细胞铺板培养5天左右，细胞融合度达到90%时可进行传代。
2.提前将培养基、PBS、胰酶放入37℃水浴锅内预热，用75%酒精擦拭后再放入生物安全柜内。
3.吸除旧培养液，加少量PBS润洗细胞，适量胰酶，使加入的胰酶能盖住细胞，37℃孵育，约2~3min显微镜下观察。
4.待贴壁细胞间间隙变大、细胞趋于圆形但还未漂起时弃去胰酶，加入新鲜培养基，晃动细胞瓶，终止胰酶作用，用吸管小心吹打贴壁的细胞，制成细胞悬液（控制吹打的力度，避免产生大量的气泡）。
5.500g，常温离心5min，去上清并用肾皮质细胞培养基重悬。
6.将细胞悬液按2×10^4cells/cm2接种到新的胶原包被培养瓶内，置37℃/5%CO2培养箱中培养，隔天观察贴壁生长情况。
7.肾皮质细胞体外传代代数≤15代，其中人肾皮质细胞代数≤9代。
The cells can be cryopreserved using standard cell line protocols.
VI Regarding After-Sales Service
Quality Assurance & After-Sales Policy In the event of product quality issues, please: Collect original data Contact our sales representatives or technical support team immediately We will dispatch personnel to address the issue promptly.
After-Sales Validity Period & Submitted Original Data：
Resuscitation issues: Report any abnormalities immediately and provide results of Trypan blue staining or AO/PI staining.
Contamination issues: Report within 96 hours after resuscitation and provide phase-contrast microscope photos.
Purity issues: Report within one month of detection and provide immunofluorescence or flow cytometry results.
VII Contact Number
Company Telephone：0755-28284050
Technical Support：19902901483（ Dr. Zhou）
