Disclosed is an industrial preparation of natural killer cells (NKs) and an injection using human allogeneic karyocytes comprising: using umbilical cord blood and peripheral blood from legitimate sources as raw materials, obtaining stem cells by a method for extracting and separating karyocytes, or using Ficoll or percoll density gradient centrifugation to isolate and screen out karyocytes diluting the above-mentioned karyocytes with cell culture medium, adding interferon, interleukin, CD3 antibody, and human albumin, loading them together into a bioreactor for perfusion culture, and then performing multiplication culture the passage number of natural killer cells from multiplication culture is no less than 8, and the culture time is no less than 4 weeks the markers of the natural killer cells obtained after the multiplication culture are CD3 - \CD56 + , CD16 + , CD57 + , and CD8 + , wherein CD16 + /CD56 + ‰¥15%, CD3 - /CD56 + ‰¥ 50%, and CD8 + /CD57 + ‰¥ 8% then preparing an injection with a certain concentration using the cell suspension obtained by above-mentioned method.