AIM: To investigate the effect of internal limiting membrane transplantation and autologous blood on treating refractory giant macular hole. METHODS: Thirty-seven eyes with giant macular hole of the smallest hole diameter >700 μm, the maximum diameter of the substrate >1000 μm and hole formation factor <0.6 underwent surgical treatment. The patients were randomly divided into two groups. Nineteen eyes with surgical flip of the internal limiting membrane in group A, 18 eyes with internal limiting membrane transplantation in group B who underwent the tamponade of internal limiting membrane into the hole, autologous plasma was used to seal the hole. The patients were followed up for 3mo, optical coherence tomography and best corrected visual acuity (BCVA) were recorded before and after operation, and the results were statistically analyzed. RESULTS: At 3mo after operation, BCVA of the two groups was significantly improved compared with that before operation (tA=4.192, tB=4.374, P<0.05). But there was no significant difference in visual acuity between the two groups (χ2=0.128, P>0.05). At 3mo after operation, the closure rate of group A was 68.4%, and 100% in group B. (χ2=5.628, P<0.05). The defect diameter of inner segment/outer segment at 3mo after the operation was significantly lower than that before operation (tA=12.287, tB=15.481, P<0.05), and the difference was statistically significant (t=2.552, P<0.05). CONCLUSION: Internal limiting membrane transplantation combined with autologous whole blood can improve the postoperative closure rate of the refractory large aperture, and can effectively improve the postoperative visual acuity.