(HUVEC) are isolated from the vein of the umbilical cord and are commonly used for physiological and pharmacological investigations, such as macromolecule transport, blood coagulation, angiogenesis, and fibrinolysis.
- 1. How can I successfully isolate HUVEC from umbilical cords without using antibiotics?
- 2. What’s the difference between PromoCell HUVECs and HUV-EC-C from ATCC?
- 3. Which type of endothelial cell is best suited for studying angiogenesis?
- 4. How many passages can I perform withPromoCell HUVECs?
- 5. Are PromoCell Cells derived from single donors or are they pooled from multiple donors? What’s the difference between them?
- 6. Which medium do PromoCell recommend to use on HUVEC – Endothelial Cell Growth Medium or Endothelial Cell Growth Medium 2?
1. How can I successfully isolate HUVEC from umbilical cords without using antibiotics?
At PromoCell, we get the umbilical cords from our tissue suppliers with no addition of buffers or media. This method prevents the microorganisms from being washed into the blood vessels. Before we start the cell preparation, the umbilical cord is also cut on both ends with a sterile scalpel to provide sterile intersections in addition to the sterile lumen.
This method allows us to isolate sterile endothelial cells from umbilical vein and to plate them in antibiotic-free culture media.
2. What’s the difference between PromoCell HUVECs and HUV-EC-C from ATCC?
PromoCell HUVECs are freshly isolated from umbilical veins. They are cryopreserved at the end of primary culture. After revival, they can be propagated for > 15 doublings and will senesce eventually.
HUV-EC-C from ATCC is a hypodiploid human cell line of endothelial origin (umbilical vein). The modal chromosome number is 45 occurring in 72% of cells counted. The rate of polyploid cells is 15.8%. The cells have a life expectancy of 50 to 60 population doublings. This indicates that the cells are no longer “normal cells” but have undergone some degree of transformation.
3. Which type of endothelial cell is best suited for studying angiogenesis?
The study of angiogenesis has been significantly advanced by the ability to culture endothelial cells in vitro. Initially, large vessel ECs, such as those isolated from the human umbilical vein (HUVEC) were used for these studies but increasingly it has been recognized that microvascular endothelial cells are a more appropriate model since angiogenesis involves microvessels rather than large vessel ECs.
4. How many passages can I perform withPromoCell HUVECs?
PromoCell guarantee > 15 PD for their HUVEC. The number of passages that can be performed, depends on the dilution factor used during subculture. If you split the cells 1:4, they can perform about 2 population doublings per passage which means that they can be cultured for at least 6-8 passages.
5. Are PromoCell Cells derived from single donors or are they pooled from multiple donors? What’s the difference between them?
PromoCell Human Cells are generally derived from single donors. The donor’s age, sex, and race is specified in the lot-specific Certificate of Analysis which can be downloaded from our website.
Our HUVEC single donor (C-12200) are isolated from a single umbilical cord, propagated in primary culture, and frozen down at subconfluency.
For the preparation of HUVEC-pooled (C12203), we simultaneously isolate the cells from 2-4 umbilical cords and grow them in separate tissue culture dishes. The cells are pooled after trypsinization given that their growth rates are comparable.
6. Which medium do PromoCell recommend to use on HUVEC – Endothelial Cell Growth Medium or Endothelial Cell Growth Medium 2?
The standard medium for isolation and propagation of our HUVEC, HUAEC, HPAEC, and HSaVEC is Endothelial Cell Growth Medium (C-22010). It contains ECGS, an extract from bovine hypothalamus which has mitogenic effects on endothelial cell proliferation.
Scientists who prefer a more defined Growth Medium can use Endothelial Cell Growth Medium 2 (C-22011). In this medium, ECGS is replaced by VEGF, IGF, and additional bFGF and EGF to stimulate endothelial cell growth.
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