60 BIOTECHNOLOGY
The collagen cell carrier F
Silke Busch looks at how collagen products produced for the food market have transferred to cell biology and biomedical applications.
or 80 years collagen type I products have been produced for the food
market in a standardised and large-scale industrial production process.
Ten the idea arose to translate this huge knowledge into the development of a special collagen membrane for use in research and development in cell biology and/or regenerative medicine, respectively tissue engineering.
Collagen is the most common protein in mammalians and is an essential structural element of the extra-cellular matrix. Tat makes collagen per se in vitro and in vivo an ideal substrate for cell and tissue culture.
Complex cell structures Tanks to its special characteristics, the collagen cell carrier (CCC) is an excellent support material for the establishment of complex cell and tissue structures (including organotypic cell culture models, like, for example, differentiated and stratified epithelial structures).
During the production process the collagen from bovine skin middle splits will be processed and ends up in a combination of an extraction and cleaning of the insoluble collagen fraction.
Fig. 1. Electron microscopy picture of the CCC. Please note, in the re-moisturised membrane you will see more fibrous structures.
www.scientistlive.com
So the collagen type I products like the CCC are neither acidic solubilised nor enzymatic digested but rather contain fibrous collagen type I. Tus the collagen mass consists of highly
concentrated collagen type I.
After intense R & D work on the collagen membrane itself (eg, membrane thickness, degree of cross-linking) and testing variations to optimise the composition of the collagen mass, first proof-of-principal studies with collagen films for cell cultivation were performed.
In these studies a lot of different adherent primary cell types were included, mesenchymal stem cells (MSC) too. Additionally, MSC can also be differentiated in all four lineages (chondrogenic, osteogenic, adipogenic and neurogenic) directly on the CCC.
Tis in vitro result together with all the analytical data shows that the knowledge transfer from production of collagen products for the food market to life science and biomedical applications is possible.
The detection line A lot of new analyses, according to ASTM 2212-02 for CCC,
were performed which shows that levels of contaminating DNA or RNA, foreign proteins, heavy metals or even endotoxins are all at or under the detection line, the collagen type I purity is 99 per cent in Mascot Score.
We have developed a new collagen type I membrane with very special characteristics:
■ Ultrathin structure with
preserved long collagen fibres; ■ No chemical cross-linking; ■ Non-porous but semi-
permeable membrane allows the exchange of soluble factors through the membrane (air- lift organotypic cell cultures
possible); ■ Cells can grow on both sides of the membrane, which makes
co-culture systems possible; ■ High cell densities can be achieved without induction of
apoptosis; ■ Cells can grow multi-layered
on top of the CCC; ■ Cell seeded CCC can be embedded for histology; both in
cryosections and also in paraffin; ■ Fixation of cells followed
Page 1 |
Page 2 |
Page 3 |
Page 4 |
Page 5 |
Page 6 |
Page 7 |
Page 8 |
Page 9 |
Page 10 |
Page 11 |
Page 12 |
Page 13 |
Page 14 |
Page 15 |
Page 16 |
Page 17 |
Page 18 |
Page 19 |
Page 20 |
Page 21 |
Page 22 |
Page 23 |
Page 24 |
Page 25 |
Page 26 |
Page 27 |
Page 28 |
Page 29 |
Page 30 |
Page 31 |
Page 32 |
Page 33 |
Page 34 |
Page 35 |
Page 36 |
Page 37 |
Page 38 |
Page 39 |
Page 40 |
Page 41 |
Page 42 |
Page 43 |
Page 44 |
Page 45 |
Page 46 |
Page 47 |
Page 48 |
Page 49 |
Page 50 |
Page 51 |
Page 52 |
Page 53 |
Page 54 |
Page 55 |
Page 56 |
Page 57 |
Page 58 |
Page 59 |
Page 60 |
Page 61 |
Page 62 |
Page 63 |
Page 64 |
Page 65 |
Page 66 |
Page 67 |
Page 68