SMC turn to Fibroblasts - (Jul/07/2015 )

Hi,

I´ve ordered SMC from human pulmonary artery. Even I´ve used the medium and all necessary supplements, ~98% of the cells didn´t attach to the flask bottom.

As the cells were super expensive I hoped for something like a miracle to happen ... so I waited 2 days until i found a few cells wich made it and attach.

Because I had nothing to lose I cultured them for more than a week and also tried to passage them which wordek fine...

Now I got some cell .... but I don´t think that they are SMC anymore. For me they look more than fibroblast.

In the end I also need fibroblast for my experiments... but are these (maybe) fibroblasts comparable with fibroblasts taken from pulmonary artery? Do they have same properties? Is this common that this happen ? 

Are these fibroblasts?

Thank you in advance,

kohlmeise

Where did you order the cells from? What medium and supplements are you using? If so few cells initially attached something was definitely wrong. If you bought them commercially you should contact the company and ask for a replacement vial. I've had no issue with this in the past from Promo/ATCC/Lonza/etc. 

With that said, in my experience growing cryopreserved primary SMCs, they can definitely look like fibroblasts during "standard" culture.

I would suggest using SMC differentiation media for a couple days and see if the morphology changes from spindle-like to rhomboid-like. You could also stain for smooth muscle alpha actin (aSMA) expression which should be increased.  

Here is SMC formulation from ATCC (really the only critical component is the heaprin though I believe): http://www.atcc.org/support/faqs/8df5c/Primary%20smooth%20muscle%20differentiation%20procedure.aspx