Purpose: The long-term development of talent in endurance sports is a topic of interest. Among various factors, the importance of total hemoglobin mass (tHbmass) and the potential benefits of being an altitude-native athlete remain unclear, particularly in young categories. This study aimed to investigate the impact of altitude and training content on hematological characteristics by comparing young male cyclists age 15–16 and 17–18 years who were born and trained at a moderate altitude (ie, greater than or equal to 2500 m; MA) and cyclists who were born and trained at low altitude (below 1000 m; LA). Methods: tHbmass (in grams and grams per kilogram), measured by using the optimized carbon monoxide rebreathing method during an incremental test on a cycle ergometer; hematocrit percentage, hemoglobin concentration; and erythrocyte, blood, and plasma volume were measured in youth male cyclists age 15–16 years and junior cyclists age 17–18 years who were born and trained at MA versus LA. All variables were analyzed with a 2-way (age [youth cyclist vs junior cyclist] × altitude level [MA vs LA]) analysis of variance with subsequent Tuckey post hoc test. Results and Conclusion: Some altitude-induced benefits were reported in cyclists at age 15–16 years in the MA group with higher values in hematocrit percentage, hemoglobin concentration, and tHbmass (grams per kilogram) (P < .05) than their LA counterparts. This was also observed at age 17–18 years (P < .001), except for tHbmass, wherein no significant difference was found between MA and LA groups. In contrast, plasma volume was lower in MA than LA junior cyclists. New Findings: (1) The altitude of birth and residence could generate an advantage in tHbmass in young male cyclists age 15–16 and 17–18 years who train at MA compared with cyclists who are born and train at LA. (2) Altitude-induced benefits in physiological variables (hematocrit percentage, hemoglobin concentration, tHbmass in g·kg−1) were reported in cyclists at age 15–16 years and partially at age 17–18 years. In contrast, plasma volume was lower in MA than in LA junior cyclists. This may impact the strategies for identifying and developing talent in cycling.