Hello Andrei, it is quite normal for cells derived from adipose tissue to be bad at growing or attaching. The reason is in the colony forming test if you use bone marrow msc's and differentiate them into  adipose cells. They really are poor at growing speed and replication. I would suggest you to change cells, or if you really want to use them, try coating the scaffolds with proteins like albumin, collagen etc. I would also have a look at deem composition and try to add some growth factors to improve the cells health

The decellularized 3D scaffolds afford the best substrate for your cells - basically collagen and other ECM proteins. If your cells attach to culture plates they should attch to 3D matrix as well.

The surfactants used in the decellularization protocol may highly influence in cell attachment and proliferaton. if you lower the surfactant concentration or increase the washing time may help to improve the cell adherence and proliferation on decellularized scaffolds. For cell seeding, try to have a high concetrated cell solution.

The scaffold should contain enough cell adhesion motifs if the blood vessel was properly decellularized.  Try to make the scaffold clean after the decellularization process.

Try dynamic seeding and may be using a higher percentage of FBS or any serum that you use in your media to condition your scaffold which might have dried out a bit form the  detergents used in decellularization, but as mentioned above your scaffold should have the BST adhesion substrate (collagen)

I am not agree with Antonio M Spadaccino, we know that one of the best charactristic of ADSC is the adhesion power. They do adhere to different ( but suitable surfaces) very nicely and fast. Did you cultured the same cells at the same time on TCPC? if they do attache to TCPC and not to your scaffolds then you need to check scaffolds. Maybe during the procedure you did something wrong and the scaffolds lost their properties. What I would suggest for loading, 

- First be sure you are loading enough healthy cells.

-pre-wash your scaffolds with your culture medium at least 3 times.

-You can clamp one site of the scaffold and load the cells in high density/volume of medium.

-Leave them for couple of hours and then start rotation. You can even load them at 2-3 steps. 

This should solve your problem.

Good luck.

Dear Andrei,

when working with 'refractory' scaffolds, we usually first treat them with 70% ethanol, which makes it more hydrophilic, then wash 2-3 times with PBS and finally 'coat' them with FBS for 3-4 hours. This would make the substrate more prone to cell attachment.

Drop-seeding of cells, that is to put on the scaffold the cells concentrated in 10-50 microliters of medium (depending on the substrate surface area), then adding the proper volume of medium after 1-2 hours, usually works. Provide high humidity to this system, otherwise the drop will dry.

Good luck!

use additional fibronectin (soak the scaffold in fibronectin solution ) if it does not interfere with your study. 

http://icvts.oxfordjournals.org/content/8/3/301.full

Passivation with serum albumin will help.....

I would say enhancing cell attachment using any of those coatings (collagen, gellatin, fibronectin, . . .) will definitely extend your study more.

You have to calculate the porosity and total volume of your scaffolds and seed your cells accordingly.

- Pre-wet your scaffolds with complete medium (trying not to fully drench them in medium) and keep them warm in the incubator shortly before seeding.

- Select the right seeding volume and cell density and seed the cells gently (in a Petri dish), not to allow the suspension to spill out of the scaffold.

- DO NOT TOP OP WITH MEDIUM IMMEDIATELY. Incubate the seeded scaffolds in the incubator immediately after seeding for at least 2 h.

- Put proper volume of pre-warmed medium (just enough to cover the scaffolds) in your well plates.

- Remove the scaffolds with small pinches using a fine tip tweezers and gently let them to sink in the wells containing medium. 

GOOD LUCK

Dear Dr Andrei Moroz

Since I face a similar problem with a decellularized biological scaffold, which of previously mentioned opinions and answers was successful?

Thanks in advance,

Hussein