3 replies to this topic

[Baraa]

Hello everyone,

I am venturing into a new field: stem cell and would like your help, please.

I isolated mouse mesenchymal stem cells (MSC) from compact bone, performed ficoll gradient centrifugation and cultured them in hypoxic conditions (5% oxygen). They have been in culture for about a week. Recently, I checked their morphology under the microscope and they seem weird to what is documented in literature. I have attached few photos. Which of these cells look like MSC?

Since cells are extracted from the compact, I am aware that there is more than one cell population isolated simultaneously. How do you proceed to obtain a homogenous MSC culture?  

Please advise. Thank you!

Attached Files

•  bioforum.pdf   604.37K   144 downloads

[bob1]

Note: I haven't done MSC culture before, but I have done a lot of other cell culture and a reasonable amount of primary culturing...

My guess is that the most likely cells to be MSC is 2, 3, 4, with #4 being the healthiest colony.  #1 appears to be some sort of fibroblastic culture, and 2 and 3 are both probably mesenchymal cells that are quite unhealthy looking (need to culture at a higher density probably) as they have long processes and indicators of attempting to migrate.

[Baraa]

Hi Bob1,
Thank you for your reply. I was wondering what makes you think the cells are unhealthy? To me, those cells look more like neurons or astrocytes with long cytoplasmic extensions.

[bob1]

Yes, similar things -neurites and dendrites are forms of processes, but you are trying to isolate mesenchymal cells - you wouldn't find neurons and astrocytes unless you used the brain...

Those long things are called processes and cells have them out either as feelers for cell:cell communication or bits left behind when the cell is moving.  A cell that is moving has two obvious features - a rounded edge with small processes which is the "leading" edge that the cell is rolling over, and a ragged "trailing" edge with bits of cytoplasm left behind as the cell moves.  Have a google search for cell migration or look here.