How to get rid of fibroblastic and contaminating cells?

Very often primary cell cultures are jeopardized by the outgrowth of contaminating fibroblasts.  The population of the target primary cells is directly affected since fibroblasts usually grow at much faster rates than other cell types.

Any solution in literature?

Melanocytes contaminated with fibroblastsA search reveals that there are numerous published efforts to inhibit fibroblast outgrowth and this process appears to be tissue specific.  For instance, the addition of melatonin to melanocytes preparations potently inhibits the proliferation of fibroblasts that were passed along in the isolation procedure from whole human skin.  Another example is that of adenosine and its inhibition of collagen and protein synthesis in cardiac fibroblasts.  Specific antibodies can act as inhibitors of fibroblasts growth and proliferation.  A DNA topoisomerase inhibitor (camptothecin) can be included in culture medium to eliminate non-neuronal cells proliferation in neuron-type primary cell isolation.  In addition, it has been reported that D-valine is a selective agent for rat epididymal epithelial cells, but not for rabbit epithelial cells, and that cytosine arabinoside is a simple and effective means to control the proliferation of fibroblast-like cells in both rat and rabbit epididymal cell cultures.

Together, a tissue-specific optimal fibroblast inhibitory system can be developed to improve the performance of primary cell isolation.

Removal of unwanted cells

FribrOut - prevent overgrowth of fibroblastic and contaminated cellsThe fibroblast growth control system, FibrOut™ developed by CHI Scientific is a proprietary combination of several biochemical compounds, antibodies, or specialty reagents, which prevent overgrowth of fibroblastic or contaminating cells, effectively increasing the yield of target cells. Each fibroblast growth inhibitory system is customized to be tissue-specific and cell-specific.

Each FibrOut™ system may contain one or more of the following chemicals (1) Trypsin; (2) Collagenase; (3) D-valine; (4) Cis-OH-proline; (5) Ethylmercurithiosalicylate; (6) Phenobarbitone, (7) Formulated serum substitutes. Some of the systems also contain antimesodermal antibodies.

Written by Jean-François Têtu, PhD
Jean-François Têtu is Sales Manager at tebu-bio for the south of Europe and also for the UK and Eire. He also fervently defends and promotes anything from his home territory, the north of France.