How do you determine what cell density could be used to suspend on a 3D PCL based scaffolds of dimensions 9 mm diameter and 3 mm thickness. Should I use a lower cell density of 0.5*106 cells/ml or a higher cell density of 4*106 cells/ml.
Thank you for your help.
This all depends on the cell type you are growing/intending to use. Smaller stem cells such as MSCs or bone cell lines (MG63 and HOSTE85) are typically smaller in size than conventional primary connective tissue cells such as fibroblasts and chondrocytes. Also, the natural cell density of the tissue you are emulating needs to be considered. It's not all about vast quantities of cells, but what quality matrix can be laid down to maintain them under the specific culture conditions. Good luck.
I asked same kind of question and I got nice feedbacks :) this is one of my questions
https://www.researchgate.net/post/What_is_the_appropriate_density_for_culturing_MSC_or_AC_on_scaffolds
Thank you for your comments.
I am using bovine chondrocytes. Some articles say that cell density should be higher for scaffolds thickness greater than 2mm which is 5-50x106 cells/ml but some articles used 0.5-1x106 cells/ml. It is confusing which protocol to follow.
Dear Dr. Jonnala,
Your questions is very interesting but to respond precisely many parameters need to be considered:
1.What type of coating are you using on your PCL fibers and what strategy are you using to coat the scaffold?
For instance in my research I used A ddH20 Fibronectin solutions(50µg/ml) as the coating solutions, the scaffolds (Collagen,Chitosan,Collagen-Chitosan) are crosslinked and whashed and than are dryed prior to coating by immersion in a solution of Fibronectin for 45 min at 37°C.In the case of PCL scaffold is recommended to O2 Plasma the dryed scaffold for 3 min prior coating to improve the coating efficacity.
2. The size and thickness of the scaffold : 3mm thick scaffold it's from my experience very thick depend of the migration/expension capacity of your cells for clinical applications the best thickness range is from 500µm-1.5 mm thick.
3.What seeding strategy are you using ?
For instance, is essential to concentrate your cells in 250µl Medium and than coat the scaffold, wait for 15-45min at 37°C and add the medium for proper bioadhesion of the cells on the scaffold.
4. Are you using a bioreactors ?
All these parameters are from my experience essential to determinate precisely the seeding density of the cells on a scaffold.
Best,
Philippe
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