A standardized protocol for the isolation and culture of normal and arthritogenic murine synovial fibroblasts
Figure 1
Figure 2
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.
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.
Posted 01 May, 2009
A standardized protocol for the isolation and culture of normal and arthritogenic murine synovial fibroblasts
Posted 01 May, 2009
Figure 1
Figure 2
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.
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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