Please find the following protocol:

Infection of Chicken Secondary Tissue Culture Cells


Overview:

This protocol describes how to virally infect a primary chicken culture.


Procedure:

1. Seed 3 X 106 Primary Chicken Cells on a 100 mm cell culture
dish in Seeding Medium.

2. Incubate at 37°C for 1 to 2 hours to allow cells to settle.

3. Replace the medium with 5 ml of Growth Medium.

4. Add 0.5 to 1.0 ml of stock virus (approximately 1 X 106
pfu/ml). Uninfected cells can be passaged in parallel as
negative control for the status of the infected culture.

5. Incubate overnight at 37°C. If the infection was successful,
the medium should acidify quite quickly (i.e. within one day).

6. Replace the Growth Medium with 10 ml of Stable Growth
Medium.

7. Incubate at 37°C and replace the Stable Growth Medium
every other day. By 4 to 5 days, infected cells should begin
rounding up. Never passage these cells at more than 1:3
dilution.


Solutions:

Stable Growth Medium...................1% DMSO in Growth Medium
................................1% Chicken Serum
Growth Medium.............................................25 ml Calf Serum
.....................................400 ml 1X 199 Medium
........................50 ml Tryptose Phosphate Broth
........................10 ml 2.8% Sodium Bicarbonate
2.8% Sodium Bicarbonate.........2.8% (w/v) Sodium Bicarbonate
Seeding Medium...................50 ml of Tryptose Phosphate Broth
.................................400 ml of 1X 199 Medium
...............................................8 ml Calf Serum


BioReagents and Chemicals

199 Medium
Calf Serum
Tryptose Phosphate Broth
Sodium Bicarbonate
Chicken Serum
DMSO


Reference Link:

http://www.bio.com/protocolstools/protocol.jhtml?id=p680