Purpose: The main purpose of the present study was to determine physiological responses in wheelchair basketball (WB) matches in relation to heart rate (HR), match load (ML), differentiated perceived exertion (dRPE), lactate concentrations (LA), and tympanic temperature (TEMP), while specifying the individual player's playing time during different types of matches. Method: Nine Spanish First Division WB players participated in the study. Three groups were determined for each observation (i.e., HR, ML, dRPE, LA, and TEMP) according to the minutes played by the WB players: players who had played 30 min to 40 min (WB30-40), those who played 20 min to 30 min, and those who played 0 min to 20 min. Results: The WB30-40 group attained statistically significantly higher results in mean HR than the other groups in league and playoff matches (p < 0.05; ?% > -25.73%; ES > -1.31, large). With regard to the difference in each variable between league and playoff matches, the differences were highlighted in the WB30-40 group according to ML (Edwards, p < 0.05, ?% = -11.14%, ES = -0.87, large; Stagno's modified TRIMP, p < 0.05, ?% = -16.95%, ES = -0.77, large). Tympanic temperature also reached higher values in playoff matches compared with league matches in WB30-40 (p < 0.05; ?% = 1.1%; ES = 0.67, moderate) but not in LA values. Conclusions: Coaches should take into account that playoff matches were more demanding than league matches, thus forcing coaches to try to peak WB players' physical performance for the former.