The invention discloses 3D bioprinting nano cellulose hydrogel ink comprising the following components, by weight, 1100 parts of nano cellulose, 900 parts of hemicellulose, 1250 parts of ultrapure water, 380 parts of cell growth factor FGF2, 175 parts of EGF, 225 parts of VEGF, 315 parts of collagen, 165 parts of fibrinogen, 60 parts of hyaluronic acid and 700 parts of ink forming agent X, a preparation method of the 3D bioprinting nano cellulose hydrogel ink includes the following steps: uniformly mixing the nanocellulose and hemicellulose and performing standing. The 3D bioprinting nano cellulose hydrogel ink has small human immunological rejection and good biodegradability, has suitable mechanical properties, and has complete functions and low price. In practical application, the stability, reliability, convenience, degradability and environmental friendliness can also meet the needs of users and bring better use prospects.