Primary cell culture from chick embryo

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BACKGROUND

Variety of cells isolated and culture directly from tissues is defined as primary culture, it contains progenitor and differentiated cells. Cells in the primary culture has a limited life span so more cells will be required for the study. Embryonic tissue is most favorite for the primary culture because cell viability is higher.

Aim: Aim of this study is to execute primary cell culture technique using chick embryo under sterile condition.

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REQUIREMENTS

Sample:     Chick embryos (12 days gestation)¹

Apparatus:    Petri dish – 60 mm

           Forceps

           Seizures

            Centrifuge tubes – 15 ml

Instrument:    Centrifuge

Chemicals:     Cell culture medium (DMEM)

            Serum and L-glutamine

Non-essential amino acids

Antibiotics/Antimycotics

Trypsin solution – 0.5%

HBSS

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PROCEDURE

To start the experiment keep the egg pointed end up and sterile with 70% ethanol further pierce shell with sterile forceps and cut the shell rounded then disentangle and decapitate the embryo and place the embryo in the petri dish and extract appendages with sterile forceps and remaining tissues should be washed with HBSS solution twice. Add trypsin solution (10 ml per each embryo) and vertex the solution at room temperature for 1 hour. Filter the mixture with sterile gauze. Centrifuge the solution for 5000 rpm for 3-5 min. Discard the supernatant and add 10 ml of culture medium and resuspend the cells in a petri dish. Incubate the plate in 37⁰C incubator with 5% CO₂ for approximately three days to attain a confluence. After monolayer formation, subculture the chick embryo cells.^2,3

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CONCLUSION

The cultured chick embryos are ready and can be used for various purposes.

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REFERENCES

1. Hamburger V, Hamilton HL. A series of normal stages in the development of the chick embryo. J Morphol. 1951;88:49–92.
   
2. Chomiak et al. Tissue culture and chicken embryo techniques for infectious laryngotracheitis virus studies. Avian diseases. 1960;4:235-46.
   
3. Thermoscientific technical note no. 29. Available at thermoscientific.com.