Cellular Reprogramming, Inc.

What do you need from us to get started?
A purchase order and some frozen fibroblasts. Send a vial containing 0.5-1M fibroblasts for each line you want reprogrammed. We require that the lines you send have been tested and found negative for mycoplasma. The fibroblasts covered by a given purchase order don't necessarily have to be batched together. You can send us the lines over a period of weeks or even months if that is more convenient, but please do alert us regarding each shipment.
What is your historical success rate at reprogramming?
Our historical success rate on low-passage (p3-p5) dermal fibroblasts is 90-100%. Please note that we cannot guarantee a successful iPSC derivation from every line submitted. In general, slow-growing, late-passage lines tend to do less well in reprogramming. While we have often been able to attain high success rates even with lower quality material, our accelerated, feeder-free reprogramming protocol is not optimized for dealing with aged or refractory fibroblast lines.
Do we have to pay for lines where the reprogramming is unsuccessful?
The 25% down payment is forfeit for non-productive fibroblast lines. The 75% balance is due for successful derivations only and is invoiced at the time of delivery.
How should the iPSCs be plated?
We recommend plating the resuspended iPSC clumps from a single cryovial to either a single 6-well format well or (preferably) divided in 1:2 ratio between two such wells. Thaw vials using standard precautions. Add 9 ml of warm DMEM/F12 dropwise to dilute the cryoprotectant while minimizing osmotic stress. Spin down for 4 minutes at 200 rcf, aspirate the supernatant and resuspend the pelleted clumps in pre-equilibrated media. Distribute to tissue culture-treated plates coated with rLN-521 or other suitable PSC substrate.
What type of culture system do you use internally for culturing the iPSCs?
We expand and stabilize iPSCs in Nutristem hPSC XF media (Biological Industries) on recombinant human Laminin-521 (BioLamina) substrate. These reagents are available directly from the manufacturers or from their distributors.
Do I have to use the Nutristem/LN-521 system for recovering and expanding the iPSCs derived from our fibroblasts?
We have customers using a variety of popular media and substrates and they report that our iPSCS usually adapt readily to different culture conditions. Nonetheless, if you are committed to an alternative culture system we recommend recovering iPSCs from thaw in the Nutristem/LN-521 system and then transitioning them into your preferred system. Alternatively, you can routinely thaw directly into your system while keeping the transitioning approach as a backup strategy to be applied with replicate vials in the event of poor recovery.
Do you provide multiple distinct clones of iPSCs for each line?
We bulk-passage colonies from reprogramming plates in order to establish p1 PSC cultures rather than using the more traditional colony-picking approach. For this reason, we do not offer multiple putative iPSC clones corresponding to each fibroblast line. The three vials of iPSCs we deliver per line are simply replicates derived from the same iPSC culture. The bulk-passaging strategy is increasingly preferred by practitioners of the inherently non-clonal mRNA reprogramming system. In contrast to the colony picking strategy, this approach allows us to take full advantage of the very high productivity often seen when reprogramming fibroblasts with mRNA. Indeed, it would be impractical to isolate individual colonies from the lawn of iPSCs that emerges in many cases. You always have the option to sub-clone from the cultures we provide if clonal analysis is critical to your application.
Do you offer cell line authentification, karyotyping, bioinformatic assays or functional characterization of the iPSCs?
We do not offer these as standard services at present. We can refer you to third party providers who can help if you are interested. We can also provide cell pellets for a small additional fee to facilitate some of these types of analysis. Please contact us to discuss your precise requirements and pricing.
Do you offer reprogramming of animal cells or other human cell types such as blood?
Currently our service is exclusively focused on human fibroblasts.