The utilization of Bi-based superconductors in magnetic field environments is limited due to their anisotropic structure and lack of efficient magnetic flux pinning sites. Therefore, to enhance the performance of Bi-based superconductors in a magnetic field, this study presents the preparation of Bi2212 high-temperature superconducting thin films with varying NiO doping levels on SrTiO3 (STO) single crystal substrates with different concentrations of NiO seed layers via the self-assembly method. Additionally, phase purity, crystal structure, surface morphology, and transport properties were thoroughly analyzed. According to the results, the NiO seed layers prepared on the STO substrates presented epitaxial growth characteristics, with the NiO second phase in the Bi2212 matrix growing epitaxially along the seed layers. As NiO doping slightly broadens the phase formation temperature range of Bi2212 superconducting thin films, the self-assembly method can be utilized to prepare NiO nanorods with epitaxial growth in Bi2212 thin films. Notably, the self-assembled NiO-doped Bi2212 thin film demonstrated higher critical current carrying capacity, with its Jc reaching 6.28 × 104 A/cm2 at 10 K in self-field. This provides a technical reference for improving the performance of high-temperature superconducting thin films under a high magnetic field.