Changing the media in which cells grow, however, is only part of the answer to providing a controlled and reproducible environment. The nature of the surface to which cells attach can also greatly affect their behavior. For example, most neural cells can not differentiate unless they are grown on a surface coated with an appropriate extracellular matrix material such as laminin. And different cells have different surface requirements. For some, simply coating the surface with positively charged material such as polylysine provides the necessary stimulation for cells to attach and prosper.
Other cell types, however, are more demanding and require complex biological coatings for successful culture. In the past, human embryonic stem cells have routinely been grown on feeder cells or surfaces coated with animal-derived extracellular matrix components. These biological surfaces have been effective in providing cells with a culture environment that mimics in vivo conditions, though their variability and propensity to contain unwanted contaminants such as growth factors has been a problem. Providing alternative growth environments for demanding cells such as these has not been easy—nevertheless, real progress is being made.
The first generation of animal-free, bio-active growth surfaces ranged from products containing relatively simple, small synthetic peptide domains to those featuring whole multi-unit proteins produced and subsequently purified from mammalian cell culture. These proteins were designed to mimic extracellular matrix components and could be coated on a variety of surfaces.
The random nature of protein attachment and presentation with first-generation products meant that only a small percentage of the active domains of these proteins were accessible and correctly displayed for interaction with cells cultured on the surface. Aside from hindering experimental efficiency, this was financially wasteful, as only a limited number of these expensive proteins was available to deliver the desired biological stimulus.
Next-generation products such as those available from Orla Protein Technologies help overcome these deficiencies. For the Orla system, a specific cell wall protein was selected and carefully engineered to auto-adhere to surfaces such as glass. It also adheres to gold, enabling parallel protein-binding studies using techniques such as SPR and QCM.
The protein has enhanced beta-barrel rigidity to aid membrane formation and upper-end peptide sequences into which peptides or protein motifs can be inserted for presentation to cells within a natural loop feature.