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Method Article
A standardized protocol for the isolation and culture of normal and arthritogenic murine synovial fibroblasts
Figure 1
Figure 2
Maria Armaka
commented on 28 April, 2014
Author's notes: 1. The protocol can be applied for the isolation of SFs from a single mouse. Optional: Volume of collagenase-digestion-solution can be adjusted from 10 to 5 ml. 2. The expected cell number stated in the protocol is the minimum one can obtain. Usually the yield of cell number per mouse, after 3 passages, is at least 4x10^7 cells (4 × 175cm^3 flasks, ~10^7 cell/flask), usually 2-2.5 weeks after isolation. 3.The suggested time-window for experiments is between 2.5-4 weeks (and always after the 3rd passage), as the cells have been shown to maintain their basic phenotypic characteristics (CD90.2+, VCAM1+, ICAM-1+) and proliferative capacity for 4 weeks.
Maria Armaka
commented on 28 April, 2014
Author's notes: 1. The protocol can be applied for the isolation of SFs from a single mouse. Optional: Volume of collagenase-digestion-solution can be adjusted from 10 to 5 ml. 2. The expected cell number stated in the protocol is the minimum one can obtain. Usually the yield of cell number per mouse, after 3 passages, is at least 4x10^7 cells (4 × 175cm^3 flasks, ~10^7 cell/flask), usually 2-2.5 weeks after isolation. 3.The suggested time-window for experiments is between 2.5-4 weeks (and always after the 3rd passage), as the cells have been shown to maintain their basic phenotypic characteristics (CD90.2+, VCAM1+, ICAM-1+) and proliferative capacity for 4 weeks.
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CURRENT STATUS: Posted
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Posted 01 May, 2009
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This is a preprint. It has not completed peer review.
Method Article
A standardized protocol for the isolation and culture of normal and arthritogenic murine synovial fibroblasts
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
History
CURRENT STATUS: Posted
Version 1
Posted 01 May, 2009
Community comments: 2
Metrics
Comments: 2
HTML Views: ...
Subject Areas
Biological techniques
Figure 1
Figure 2
Maria Armaka
commented on 28 April, 2014
Author's notes: 1. The protocol can be applied for the isolation of SFs from a single mouse. Optional: Volume of collagenase-digestion-solution can be adjusted from 10 to 5 ml. 2. The expected cell number stated in the protocol is the minimum one can obtain. Usually the yield of cell number per mouse, after 3 passages, is at least 4x10^7 cells (4 × 175cm^3 flasks, ~10^7 cell/flask), usually 2-2.5 weeks after isolation. 3.The suggested time-window for experiments is between 2.5-4 weeks (and always after the 3rd passage), as the cells have been shown to maintain their basic phenotypic characteristics (CD90.2+, VCAM1+, ICAM-1+) and proliferative capacity for 4 weeks.
Maria Armaka
commented on 28 April, 2014
Author's notes: 1. The protocol can be applied for the isolation of SFs from a single mouse. Optional: Volume of collagenase-digestion-solution can be adjusted from 10 to 5 ml. 2. The expected cell number stated in the protocol is the minimum one can obtain. Usually the yield of cell number per mouse, after 3 passages, is at least 4x10^7 cells (4 × 175cm^3 flasks, ~10^7 cell/flask), usually 2-2.5 weeks after isolation. 3.The suggested time-window for experiments is between 2.5-4 weeks (and always after the 3rd passage), as the cells have been shown to maintain their basic phenotypic characteristics (CD90.2+, VCAM1+, ICAM-1+) and proliferative capacity for 4 weeks.