In Vitro Stimulation of RAW264.7 Macrophages

2024-09-06

In Vitro Stimulation of RAW264.7 Macrophages

Goals:

Culture RAW264.7 macrophages in vitro.

Perform in vitro stimulation with LPS and measure cytokine production, specifically TNF-α using ELISA.

RAW264.7 (ATCC® TIB-71™) Macrophage Cell Line:

is derived from murine macrophages, immortalized through viral transformation or cell fusion.

is used to study macrophage behavior, immune responses, and pathogen interaction in vitro.

Macrophages serve as phagocytes, removing pathogens and signaling other immune cells via cytokines and chemokines.

The microscopy of image of the counting chamber

Trypan Blue is a dead cell exclusion dye, distinguishing live cells (unstained) from dead cells (blue). Use a hemocytometer to count live and dead cells from the Trypan Blue mixture.

Number of living cells: 64

dilution factor: 2

Total of 20 mL = 7.8125 mL from a cell culture dish with pre-seeded RAW264.7 cells + 12.1875 mL of DMEM

Macrophages are observed via microscopy.

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