Isolation and Maintenance of Endothelial Stem Cell - Creative Biolabs

Creative Biolabs is dedicated to providing the latest and most innovative protocols for stem cell study. Here we will be discussing the isolation and maintenance of endothelial stem cells. Endothelial stem cells are an area of research interest because of their potential to regenerate blood vessels and restore blood flow to damaged tissues.

The isolation and maintenance of endothelial stem cells is a complex and multifaceted process, requiring careful attention to detail and precise handling techniques. We describe a protocol for the isolation and maintenance of endothelial stem cells.

Materials Required

• Tissue source
• Medium
• Fetal bovine serum
• Endothelial growth factors
• Antibiotics
• Trypsin-EDTA
• Cell culture flasks and plates

Procedure

Isolation

Wash the tissue thoroughly with PBS to remove any debris or blood. And use enzymatic digestion or mechanical disruption to isolate the endothelial stem cells. Then collect the cells and centrifuge to form a pellet.Discard the supernatant and resuspend the pellet.

Culture and Expansion

Plate the cells in flask and incubate. After 24 hours, replace the media with fresh medium containing 10% FBS and endothelial growth factors. Monitor the cells closely for signs of growth and proliferation. Replace the media every 2-3 days. Repeat the above steps until the desired cell number is obtained.

Maintenance

The cells can be maintained in culture or frozen for future use. For maintenance in culture, subculture the cells every 3-4 days to maintain their viability and proliferation.Replace the media every 2-3 days with fresh medium.

Notes

• Always work under sterile conditions to avoid contamination.
• Be careful not to over-trypsinize the cells, as this can lead to their death.
• It is important to optimize the culture conditions and growth factors to ensure optimal growth and proliferation of the cells.
• It is critical to monitor the cells closely for any changes in morphology or gene expression, as this can provide valuable insights into their behavior and potential applications.

At Creative Biolabs, we are dedicated to providing the highest quality protocols and materials to ensure the success of your experiments. We hope that this protocol has been informative and helpful in your research endeavors.

Reference

1. Ingram D A, et al. Identification of a novel hierarchy of endothelial progenitor cells using human peripheral and umbilical cord blood. Blood, 2004, 104(9), 2752-2760.

For Research Use Only. Not For Clinical Use.