As stated in the question, I would like to know what kind of culturing medium is generally used for culturing primary human VSMC.
Currently for murine cells I'm using DMEM with 10% FBS, Pen/Strep, GlutaMax, non-essential aminoacids and DNase type I. I'm doubting if this type of medium will work optimally for human derived VSMCs. I've read something about M199 suppelemented with 10% FCS and EGF.
https://www.ncbi.nlm.nih.gov/pubmed/22057457
https://www.ncbi.nlm.nih.gov/pubmed/22222531
https://www.ncbi.nlm.nih.gov/pubmed/19301668
Dear Chandra, thank you for the reply.
I've had a look at the links you recommended. The first paper/protocol is where they mention the M199 medium in their methods. Unfortunately I couldn't access the 2nd paper. The third paper doesn't mention human VSMCs, only murine.
Do you happen to have any experience in culturing VSMCs with M199 medium (or any other kind of media for that matter)?
Hi Margit,
Thank you very much for your detailed answer! (and for the fact cells can get fat, didn't know that ;))
The problem is our cell culture is very limited, since these cells were obtained directly from a patient. I have ordered M199 medium and I will check in parallel which medium would work best for me.
Thanks!
Hi Maurice,
I have used both M199 and DMEM or DMEM+HamF12 (SFM for 48 hours).
Most of our publications in VSMC are from rabbit, but I have cultured human VSMC too in the same medium, but that was over 20 years back.
All my publications in VSMC are listed in my RG page,
just giving you 2 below, one is in JBC available free online, and full text of these 2 papers are available here.
file:///C:/Documents%20and%20Settings/farina/My%20Documents/Downloads/SomasundaramC_JBC_1995%20(1).pdf
https://www.researchgate.net/publication/14663851_A_novel_smooth_muscle-specific_enhancer_regulates_transcription_of_the_smooth_muscle_myosin_heavy_chain_gene_in_vascular_smooth_muscle_cells
https://www.researchgate.net/publication/269634857_Regulation_of_vascular_smooth_muscle_myosin_heavy_chain_gene_expression_in_cultured_vascular_smooth_muscle_cells_by_growth_factors_and_contractile_agonists
Article A novel smooth muscle-specific enhancer regulates transcript...
Article Regulation of vascular smooth muscle myosin heavy chain gene...
Thanks for the suggestions.
I took a shot and kept culturing with the medium I described in the question (DMEM with FBS, GlutaMAX, Pen/Strep, NEAA and DNase I) and it worked out fine for human derived VSMCs. For an existing culture of murine VSMCs I switched to growing them in M199 medium, but this didn't seem to have a huge effect on cell growth.
- Badges
- Science topic
- Similar topics
- Biological Science
- Cell Biology