Primary microglia were isolated according to the method of Giulian and Baker (1986) (link). After removing the meninges, cells were separated using mechanical shearing and 0,25% trypsin (GIBCO by lifeTechnologies™). Subsequently, cells were transferred into Poly-L-Lysine (PLL) (Sigma-Aldrich) coated T75 culture flasks (Greiner Bio-One) and cultured under standard conditions at 37°C and 5% CO2 (1-2 brains per flask). On the next day, flasks were washed three times with Dulbecco’s Phosphate Buffered Saline (DPBS) (GIBCO) and cultured for an additional 7-10 days in Dulbecco’s modified Eagle’s medium (DMEM) (GIBCO) containing 10% heat-inactivated Fetal Bovine Serum (iFBS) (GIBCO), 1% Penicillin/Streptomycin (P/S) (GIBCO) and 1 mL of filtered L929 cell supernatant as a source for growth factors. Cultures were regularly checked for loosely attached mature microglia. Finally, microglia were shaken off from the astrocyte monolayer after 7-10 days followed by two more shake off cycles every second to third day.
Fetal bovine serum ifbs
Manufactured by Thermo Fisher Scientific
Fetal Bovine Serum (iFBS) is a cell culture supplement derived from the blood of bovine fetuses. It provides a rich source of proteins, growth factors, and other nutrients essential for the growth and maintenance of various cell types in vitro.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using fetal bovine serum ifbs
1
Isolation of Primary Microglia
2
Isolation of Primary Microglia
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primary microglia were isolated according to the method of Giulian and Baker (1986) (link). After removing the meninges, cells were separated using mechanical shearing and 0,25% trypsin (GIBCO by lifeTechnologies™). Subsequently, cells were transferred into Poly-L-Lysine (PLL) (Sigma-Aldrich) coated T75 culture flasks (Greiner Bio-One) and cultured under standard conditions at 37°C and 5% CO2 (1-2 brains per flask). On the next day, flasks were washed three times with Dulbecco’s Phosphate Buffered Saline (DPBS) (GIBCO) and cultured for an additional 7-10 days in Dulbecco’s modified Eagle’s medium (DMEM) (GIBCO) containing 10% heat-inactivated Fetal Bovine Serum (iFBS) (GIBCO), 1% Penicillin/Streptomycin (P/S) (GIBCO) and 1 mL of filtered L929 cell supernatant as a source for growth factors. Cultures were regularly checked for loosely attached mature microglia. Finally, microglia were shaken off from the astrocyte monolayer after 7-10 days followed by two more shake off cycles every second to third day.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!