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Sperm cryopreservation is an effective fertility preservation method for cancer patients before anticancer treatments. However, there are little data on fertility preservation in large cohorts of patients with cancer in southern China. This retrospective cross-sectional study aimed to assess the fertility preservation status of 1034 newly diagnosed male patients with cancer in the Human Sperm Bank of Guangdong Province in southern China (Guangzhou, China). Of these, 302 patients had reproductive system tumors, mostly testicular cancers (99.0%), and 732 had other tumors, including lymphoma (33.1%), gastrointestinal cancer (16.3%), nasopharyngeal carcinoma (15.7%), leukemia (7.7%), sarcoma (3.6%), and others (23.6%). Patients with reproductive system tumors had lower sperm concentration and prefreezing and post-thawing progressive motility than those with non-reproductive system tumors (all P < 0.001). Differences in sperm concentration, progressive motility, and normal morphology rate were observed between patients with and without anticancer surgery before sperm cryopreservation (all P < 0.05). As of April 30, 2022, 63 patients used their cryopreserved sperm for assisted reproductive technology treatments and 39 pregnancies were achieved. This study provides valuable data on the fertility preservation status in newly diagnosed cancer patients in southern China, demonstrating that patients with reproductive system tumors had poor sperm quality for their pretreatment fertility preservation.