This systematic review aims to compare and evaluate the outcome of using either testicular sperm (Testi-ICSI) or ejaculated sperm (Ejac-ICSI) in intracytoplasmic sperm injections in patients with recurrent ICSI failure. The Cochrane Library, MEDLINE, EMBASE, and PubMed were used to search for relevant papers up till October 2020. Four cohort studies and two case series studies were included. Four studies investigated males with high sperm DNA fragmentation (SDF) and were classified as ''high SDF'', which included 247 couples and 2712 injected oocytes. The other three studies provided paired data to an unselected population of infertile men with either untested SDF or when anomalous SDF was not used as the basis for deciding to use Testi-ICSI, and were classified as ''noclassify'' in this study. This subgroup consisted of a total of 290 couples and 1061 injected oocytes. There was a higher level of clinical pregnancy rates (CPRs) in the ''high SDF'' subgroup when Testi-ICSI was used as compared to Ejac-ICSI, at 43.4% and 20.8% respectively, with a pooled odds ratio (OR) of 2.87 (95% confidence interval (CI) 1.44–5.71; P = 0.003). Furthermore, in the ''high SDF'' subgroup, Testi-ICSI use was associated with better take home baby rates (38%) as compared to Ejac-ICSI (16%), with a pooled OR of 3.24 (95% CI 1.20–8.76; P = 0.02). In the ''noclassify'' group, there was no statistically significant difference in the CPRs and take home baby rates of Testi-ICSI and Ejac-ICSI, although there was a trend of better CPRs and take home baby rates with Testi-ICSI use. Utilization of Testi-ICSI in recurrent ICSI failure couples, where males were confirmed to have high SDF in their ejaculated sperm, were correlated with greater CPRs and take home baby rates. However, Testi-ICSI may not result in better ICSI outcomes among men with untested SDF or when anomalous SDF was not the main factor influencing the decision to utilize Testi-ICSI.