Esch (esch at aol.com) wrote:
: I am investigating cell culture systems to propagate human diploid cell
: strain WI-38 or similar strains as a substrate for virus production. I
: need a system that maximizes surface area for cell attachment and allows
: for sampling, inoculation and easy harvesting. Seeking advise and
: recommendations for small and large scale systems.
: Thanks BobEsch
I propagatedhuman embryonnic cells as WI-38, KMS and MRC-5. WI-38 are the
easiest to propagate. I used DMEM 20% SVF with non essential amino-acids
and vitamins. Cell culture were made in 150 mm dishes. subcultures were
processed just before confluence to avoid long time exposure to trypsin.
You have to check all kinds of dishes you can buy. For my purpose, only
Falcon dishes were accepted by these cells. Their doubling tima was
approximatively around 24 hours for WI-38 and KMS.
I used these cells to determine their permissivity to parvoviruses (H-1 and
MVMp), the same cells transformed by physical or viral agents were used too
(WI38CT-1, KMST-6 and MRC5-V1.
These cells (parental cells) are not easy to cultivate. I split them 1/3
with 30% of freshly conditionned medium and 70% fresh medium.
Paul Becquart
Institut Pasteur de Lille
Virology group - Oncologie moleculaire
1 rue calmette
BP 245
59019 Lille Cedex
France
e-mail: becquart at infobiogen.fr
