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Introduction of DT40 cells into chick embryos
1Division of Bioresource and Bioenvironmental Sciences, Graduate School Kyushu University, Fukuoka, Japan.
Aim:
To examine the transfection of exogenous genes into chick embryos, applying the characteristics of avian leukosis virus (ALV)-induced chicken B cell line DT40 to the production of chimeric birds.
Methods:
The DT40 cells incorporated with exogenous gene ( lacZ constructs encoding Escherichia coli beta-galactosidase: beta-gal) were introduced into chick embryos by the injection of cells into stage X blastoderm. Manipulated eggs were incubated for 3 (trial 1) or 6 (trial 2) days, and the expression of lacZ DNA was detected by a histochemical staining method of beta-galactosidase and polymerase chain reaction (PCR) analysis.
Results:
The survival rates of the manipulated embryos incubated for 3 days (stage 18-20: trial 1) and 6 days (stage 28, 30: trial 2) were about 42% and 38%, respectively. The expression rates of the lacZ gene in the embryos in the trials 1 and 2 were about 60% and 23%, respectively, for the survived embryos.
Conclusion:
The rate of embryonic viability and expression rate of introduced genes were not so high, but it suggested the possibility of utilizing the DT40 cells as a vector for carrying exogenous genes into chick embryos.