Sep 11, 2020

Public workspaceFeeding T75, T150, and 6wp Backup

DOI
dx.doi.org/10.17504/protocols.io.8gkhtuw
  • 1University of California, San Francisco
  • Neurodegeneration Method Development Community
    Tech. support email: ndcn-help@chanzuckerberg.com
Andrea Argouarch
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DOI: dx.doi.org/10.17504/protocols.io.8gkhtuw
Protocol Citation: Andrea Argouarch 2020. Feeding T75, T150, and 6wp Backup. protocols.io https://dx.doi.org/10.17504/protocols.io.8gkhtuw
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: October 19, 2019
Last Modified: September 11, 2020
Protocol Integer ID: 28908
Abstract
Protocol includes feeding of T75 flask, T150 flask, or the back up 6 well plate.
Materials
STEP MATERIALS
ReagentFetal Bovine Serum VWR International (Avantor)Catalog #97068-091
ReagentDMEM, high glucose, pyruvateThermo FisherCatalog #11995073
ReagentPenicillin-StreptomycinGibco - Thermo Fisher ScientificCatalog #15140122
Protocol materials
ReagentFetal Bovine Serum VWR International (Avantor)Catalog #97068-091
ReagentDMEM, high glucose, pyruvateThermo FisherCatalog #11995073
ReagentPenicillin-StreptomycinGibco - Thermo Fisher ScientificCatalog #15140122
ReagentDMEM, high glucose, pyruvateThermo FisherCatalog #11995073
ReagentPenicillin-StreptomycinGibco - Thermo Fisher ScientificCatalog #15140122
ReagentFetal Bovine Serum VWR International (Avantor)Catalog #97068-091
Observations
Observations
Observe cell morphology and outgrowth
Feed every 2-3 days until 90-100% confluent
Preparation
Preparation
Turn off UV lights and clean hood with 70% ethanol
Clean items with 70% ethanol and bring into hood
a. Sterile Filtered Media with PenStrep

ReagentDMEM, high glucose, pyruvateThermo FisherCatalog #11995073

ReagentPenicillin-StreptomycinGibco - Thermo Fisher ScientificCatalog #15140122

ReagentFetal Bovine Serum VWR International (Avantor)Catalog #97068-091

Write down observations on confluency
Culturing
Culturing
Aspirate old media
Add 20 mls Amount20 mL of media with penstrep to T75 flask

Add 30 mls Amount30 mL of media with penstrep to T150 flask

Add 2 mls Amount2 mL of media with penstrep to 6wp backup

Continue to feed every 2-3 days until 90-100% confluent
Clean Up
Clean Up
Throw away biohazard materials properly
Clean and sterilize hood with 70% ethanol and turn on UV
Update cell culture notes in lab notebook