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Some of our customers have successfully used PromoCell Endothelial Cell Growth Medium MV2 (C-22022) when co-culturing Human Coronary Artery Endothelial Cells (HCAEC; C-12221) and Normal Human Dermal Fibroblasts (NHDF; C-12300).
Both cytokines are produced in E.coli.
No; we don't perform CD90 immunomagnetic separation with our pericytes. We can however exclude fibroblast contamination as follows:
1) The presence of fibroblasts in our pericyte cultures would be detectable shortly after cell isolation; as pericytes need up to 2 weeks before they start proliferating. Fibroblasts on the other hand would proliferate immediately and overgrow the culture.
2) Characterization of the isolated pericytes during QC includes flow cytometry of CD146. Pericytes express CD146 whereas placental fibroblasts don't.
Our HAoAF (C-12380) are isolated from the Adventitia the outer layer of the aorta. The cells have been characterized as fibroblasts by the expression of fibroblast-specific CD90.
The PromoCell quality control procedure includes the cultivation of fibroblasts for 15 population doublings. This is to ensure that the cells can be grown for a minimum of 15 PDs (6-8 passages depending on the split ratio used) but it does not mean that the cells immediately senesce after that point. We don't determine the maximum number of doublings or passages but most NHDF lots will certainly achieve > 20 passages.