The invention discloses an enclosed bioreactor. The enclosed bioreactor comprises a solution container, a collection container, a bioreaction tank, a liquid inlet pipeline and a liquid outlet pipeline, wherein the liquid inlet pipeline and the liquid outlet pipeline are provided with one four-way joint respectively. The invention further discloses a cell culturing method of the enclosed bioreactor. The cell culturing method comprises pre-culturing, bioreactor cell inoculation, bioreactor cell perfusion culture, bioreactor virus vaccination, bioreactor washing and replacement as well as bioreactor virus perfusion culture. By adopting the enclosed bioreactor, bioreactor perfusion culture can be realized, rapid liquid exchanging is realized, high-pressure cleaning is facilitated, repeated used is realized, the production cost is reduced, the pollution risk can be lowered, and the working efficiency is improved. By adopting the enclosed bioreactor and the cell culturing method thereof disclosed by the invention, the production operation of the bioreactor with a single pipeline in the existing method is substituted, and the aseptic requirement in vaccine production is effectively ensured. The enclosed bioreactor is easy and convenient to operate, and is suitable for fixed bed bioreactor large-scale culture.