Employing Reporter Cells in a Co-Culture to Study Retinoic Acid Production by Mouse Embryonic Cells
Transcript
Take a mouse embryo at an early developmental stage immersed in a buffer. Transfer the embryo into an enzyme solution and incubate it under physiological conditions, allowing the enzymes to break down the intercellular connections.
Mechanically dissociate the digested embryo to form a cell suspension.
Transfer the embryonic cells onto an adherent culture of reporter cells.
The reporter cells carry a transgenic construct containing the lacZ gene, which encodes the enzyme beta-galactosidase. The expression of lacZ is regulated by a retinoic acid response element, or RARE.
The embryonic cells release retinoic acid, or RA, a signaling molecule for embryonic development.
RA enters the reporter cells and binds to its receptor complex on the RARE, triggering the formation of a transcriptional activator complex.
The complex activates the expression of lacZ, resulting in the production of beta-galactosidase.
Beta-galactosidase production in the reporter cells indicates the release of RA by the embryonic cells.
