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Porcine Cryopreserved Cell and culture medium

Endothelial Cell Culture Kit

Background

Endothelial cells of ENC01 are isolated from porcine aorta. They are characterized by acetylated LDL uptake. The kit contains cryopreserved endothelial cells and culture medium.

Kit component

Components/Storage
Components Size Quantity Storage Conditions Shelf Life
Endothelial Cells, Porcine 5 x 105 cells/vial 1 Liquid Nitrogen 6 months
Culture Medium (Code: PMC-ENCM-COS) 250 ml 1 -20°C Freezer 6 months

*ENCM is a complete medium designed for optimal culture of porcine endothelial cells in vitro. They are sterile, liquid basal medium (DMEM) which contain essential and non-essential amino acids, vitamins, other organic compounds, trace minerals, inorganic salts, FBS, and antibiotics.


Precautions 
・Read the instructions carefully before beginning the culture.
・This kit is for research use only, not for human or diagnostic use.
・Always wear gloves and lab coat when handling the cell culture.

 

Protocol

  1. Thaw the Culture Medium (Code: PMC-ENCM-COS) in a 37°C water bath with gentle shaking.
  2. Add 10 mL of Culture Medium into a 15ml centrifuge tube.
  3. Quickly thaw the endothelial cell vial in a 37°C water bath. (for 1min 15sec)
  4. Transfer thawed cells into a 15 ml centrifuge tube containing 10 ml of Culture Medium and mix gently. Centrifuge for 5 minutes at 4°C at 200 g.
  5. After removing the supernatant, resuspend cells in 10 ml of the medium.
  6. Centrifuge for 5 minutes at 4°C at 200 g. After removing the supernatant, resuspend cells in 10 ml of the medium.
  7. Dispense 5 ml of cell suspension to 25 cm2 fl asks. If seeding into well plates or dishes, the recommended density is 1.0-1.5 x 104 cells / cm2.
  8. Incubate the plate at 37°C under 5% CO2, 100% humidity for overnight.
  9. Change 5 ml of Culture Medium the day after seeding and every other day thereafter.
  10. Endothelial cell culture becomes 80% within 2-3 days.

Subculture

  1. Subculture the cells when they are 80% confl uent.
  2. Rinse twice the cells with room temperature Hank’s BSS or PBS.
  3. Replace 3 ml of trypsin/EDTA solution (0.05% trypsin, 0.53mM EDTA) into fl ask and remove it immediately.
  4. Cells are covered by trypsin/EDTA solution; continue trypsinization until cells are completely rounded up (monitored with inverted microscope).
    Caution: If only a few cells detach, warm the fl ask at 37°C.We do not recommend continueing over 15 minutes of trypsinization.
  5. Add Culture Medium to stop of the trypsinization, and then transfer the cells into a centrifuge tube. Centrifuge for 5 minutes at 4°C at 200 g.
  6. After removing the supernatant, add culture medium. Count cell and plate cells in a new culture vessel.

Product List

Product Name Cat# Quantity Price

Endothelial Cell Culture Kit V-1

PMC-ENC01-COS 1SET

¥ 60,000
$ 800
€ 600

Endothelial Cells Culture Medium

PMC-ENCM-COS 500ML

¥ 27,500
$ 367
€ 275

To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.