Feeding cells

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Feeder cells (Engl. Feeder cells , feeder layer ) are used in cell cultures of the diet of cells that for later. B. to be isolated as clones or in the case of cells with a small number that grow better in a higher cell density.

properties

Feeding cells are usually somatic cells, for example fibroblasts obtained from connective tissue (e.g. mouse embryonic fibroblasts , MEF cells ) that have lost their ability to divide through radioactive irradiation or treatment with mitomycin C. This allows a fed cell to be isolated in clonal purity without isolating other growing cell types, e.g. B. in the generation of hybridoma or induced pluripotent stem cells. The fibroblasts also serve for better attachment of the cells to be cultivated. In particular, poorly growing cell cultures benefit from this. Furthermore, both cell growth and the avoidance of an anoikis depend on the cell density, which is why a minimum cell density may be necessary in the case of fed cells, which are only present in small numbers, by adding feeder cells. However, since feeder cells pose a contamination risk, approaches are being investigated to replace the use of feeder cells by adding conditioned medium, basement membrane-like matrices and / or recombinant components of the extracellular matrix. A nutrient solution in which these components have been enriched by incubation with cells is also referred to as a conditioned medium .

The feeder cells have a short lifespan due to irradiation or the addition of cell division inhibitors (e.g. mitomycin C ) and produce survival signals from the extracellular matrix as well as cytokines and growth factors that can induce embryonic stem cells to differentiate . Due to the limited lifespan and the lack of the possibility of cell division, the feeder cells die after a few days to weeks and the now increased number of the fed cells can be isolated from a single cell type .

Individual evidence

  1. ↑ NIH definition of 'feeder layer'. Retrieved June 17, 2013.
  2. ↑ NIH on the use of a feeder layer. ( Memento of the original from August 31, 2016 in the Internet Archive ) Info: The archive link was inserted automatically and has not yet been checked. Please check the original and archive link according to the instructions and then remove this notice. Retrieved June 17, 2013. @1@ 2Template: Webachiv / IABot / stemcells.nih.gov
  3. ↑ Sabine Schmitz: Der Experimentator: Zellkultur Spektrum Akademischer Verlag, 2011, ISBN 9783827425720 , p. 84
  4. ↑ A. Nagy, M. Gertsenstein, K. Vintersten, R. Behringer: Preparing Feeder Cell Layers from STO or Mouse Embryo Fibroblast (MEF) Cells: Treatment with Mitomycin C. In: CSH protocols. Volume 2006, Number 1, 2006, S., doi : 10.1101 / pdb.prot4399 , PMID 22485759 .
  5. ↑ C. Unger, H. Skottman, P. Blomberg, MS Dilber, O. Hovatta: Good manufacturing practice and clinical-grade human embryonic stem cell lines. In: Hum Mol Genet. (2008), Volume 17 (R1), pp. R48-53. PMID 18632697 . PDF .
  6. ^ RM Roberts, SJ Fisher: Trophoblast stem cells. In: Biol Reprod. (2011), Volume 84 (3), pp. 412-21. PMID 21106963 ; PMC 3043125 (free full text).