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Can you retrieve the cells from the RAFT Collagen Matrix by dissociation?

It is possible to dissociate the cells from the collagen matrix, by treating them with collagenase I or IV, both of which cleave the bond between a neutral amino acid (X) and glycine in the sequence Pro-X-Gly-Pro, which is found with high frequency...

Can I change the concentration of collagen? How thick or thin can the RAFT System get the collagen layer?

The standard RAFT™ Protocol yields a culture of 100-120µm thickness at a collagen concentration of about 80mg/mL. It is possible to increase the collagen concentration to 100mg/mL by leaving the RAFT™ absorber in contact with the hydrogel for 30...

What is the component labeled "MCGS" in the mesenchymal stem cell growth and differentiation media?

Mesenchymal Cell Growth Supplement (MCGS) is 100% fetal bovine serum prescreened to neither induce nor prevent differentiation of human bone marrow derived stem cells PT-2501

Can I add additional matrix proteins such as laminin and collagen type IV into a RAFT Culture with the addition of some basement membrane extract such as Matrigel or Geltrex?

We have tested adding up to 5%v/v Matrigel™ into the RAFT™ Collagen Mixture. It should be noted that this requires that the absorbers be left in contact with the hydrogel for 30 minutes instead of 15 minutes (step 5 of the standard protocol). All...

What is the difference between the various mesenchymal stem cell growth and differentiation basal media?

The basal media portions of the MSCGM growth and differentiation media are exactly the same except for the volume and the concentration of glucose. The glucose concentration in the growth (PT-3238) and osteogenic differentiation (PT-3924) basal media...

Can I add an additional layer of matrix proteins e.g. laminin or fibronectin at the end of the RAFT Process?

It is possible to add a layer of laminin or fibronectin at the end of an experiment after the RAFT™ Process has taken place. However, if cells have been embedded in the RAFT™ Culture, it is important to make sure that the culture is not left to dry,...

How can I create co-cultures using the RAFT3D Cell Culture System?

You can create co-cultures in different ways, which depend on the structure of the model you are trying to make. Below are three examples of co-cultures: 1. Two cell types mixed together inside one RAFT™ Culture: For this, it is possible to mix...

How does the in-vitro fm data predict the in-vivo fm value?

The Silensomes™ HLM do not need to incorporate a Relative Activity Factor and therefore directly predict the CYP contribution to drug metabolism. In-vitro fm values calculated using Silensomes™ HLM are directly correlated to in-vivo fm values using...

What is the intra-plate and inter-plate reproducibility of the RAFT System?

In terms of culture thickness, the intraplate variability is 10%; while the interplate variability is 8% (data obtained from measurements on 107 separate 96-well or 24-well insert plates).

When should the in-vitro fm be calculated in the course of drug development?

The in-vitro fm can be calculated: - early in the development of the test compound, by using the Silensomes™ HLM of 3 key CYPs involved in the biotransformation of many drugs, CYP3A4 and CYP1A2, and CYP2D6. (Catalog numbers SIL200K, SIL210K, and...
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