In the present study, we designed an ultrasensitive sensing platform for the evaluation of the physiologically relevant values of basal dopamine (DA) in a culture medium as a complex biological environment. The proposed sensing platform was fabricated via the integration of molecular imprinting technology with carbon hybrid nanomaterials. Carbon nanofibers (CNFs) were grown by using plasma-enhanced chemical vapor deposition (PECVD) on tetrahedral amorphous carbon (ta-C) thin films on silicon wafers. The prepared ta-C/CNFs sensing platforms were electrochemically coated with DA-imprinted polypyrrole as the molecularly imprinted polymer (MIP) or "artificial receptors". The three-dimensional MIP receptors were able to determine trace values of DA in phosphate-buffered saline solution (PBS) pH 7.4 (LOD = 5.43 nM) as well as in the absolute culture media such as DMEM/F-12 medium (LOD = 39 nM), DMEM/F-12 medium supplemented with 15% horse serum and 2.5% fetal bovine serum (LOD = 53.26 nM), and F-12 K cell culture medium (LOD = 62.57 nM), with highly physiologically relevant sensitivity and free of interference by other coexisting biomolecules and biological compounds. As all the fabrication steps of the composite electrode are compatible with common microsystem technology processes, the present results pave the way for integrating these ultra-sensitive electrodes to microelectrode arrays (MEA) platforms used for human dopaminergic neurons studies in vitro and enable continuous measurement of the basal DA concentration in real-time for instance in organoid studies.