Hello everyone,
I just started working with the HCASMCs which purchased from Promocell. I thaw and plate the cell in T75 flask. And used the smooth muscle cell growth media 2 (PromoCell) for culture. However, the cells grow very slowly (over 7 days but didn't reach confluency yet, maybe around 50% only ). So does anyone know how many days it takes to reach confluency? or what kind of supplement factor I can add?
Hi Yu An Lu,
It depends on the cell lot (each lot is from a different donor and some proliferate faster than others), but in general, the HCASMC have population doubling time of ~ 28-80 hrs.
I have to admit that I don't know exactly, how long it will take to reach confluence in a T75 flask - assuming that you seeded them at the recommended 7,500 - 10,000 cells per cm² , I guess it will take about 6 to 9 days.
You can check this in the CoA of your cell lot:
https://www.promocell.com/certificate-of-analysis/
As growth medium, you are using the Smooth Muscle Cell Growth Medium 2, meaning the basal media plus the included supplement mix, right? That's all the cells need for an optimal growth.
Maybe something is slowing down your cells. You are adding antibiotics? A slower proliferation is a side effect of them:
https://www.promocell.com/in-the-lab/antibiotics-in-cell-culture-friend-or-enemy/
Please feel free to get in touch with our technical support - they will be happy to look into this together with you - you can find their contact details here:
https://www.promocell.com/about-us/contact/
Hope this helps.
Kind regards,
Alexander
The PromoCell is a good culture medium for human vascular cells, but I think we should first use Flasks of 25 cm2 to give strength to the cells to proliferate after you use the flasks of 75 cm2
Alexander Trampe, Thanks for your useful information.
These days we have some issues with the Smooth Muscle Cell Growth Medium 2(C-22062). I hope to know whether mix the smooth muscle cell basal media 2 (c-22262) with the Smooth Muscle Cell Growth Medium 2 supplement Mix (c-39267) can be used as a culture medium for the HCASMCs? In other words, is it equal to Smooth Muscle Cell Growth Medium 2 (C-22062)?
If you know any information about this issue, please kindly help us to solve the problems. Thanks.
Yes, the Smooth Muscle Cell Growth Medium 2 (Ready-to-use, C-22062) consists of 1x 500ml Basal Medium (cat. #C-22262) plus SupplementMix (1 Mix for 500 ml; cat. #C-39267). It's cheaper to order the Ready-To-Use Kit than the individual components.
Oh, and to prevent any confusion: "ready-to-use" refers here to the pre-mixed supplements. You still need to add the supplement mix to the basal medium, of course.
And my apologies for the late reply, Yu An Lu! Please feel free to contact us directly to get a faster answer:
https://www.promocell.com/services/scientific-support/
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