Healthy seed cane supply is the lifeline in the productivity and profitability of sugarcane and hence the sustainability of the sugar industry, the Ethiopian sugar estates/projects have been using tissue cultured sugarcane planting materials since 2011/12. However, there is no study that shows the practical advantage of tissue culture over the conventional seed sources at Ethiopian situations. Thus, the current work was carried out to evaluate the two seed sources of two sugarcane genotypes. Accordingly, analysis of variance proved that the interaction effects of genotype by seed source is highly significant (p<0.001 at α=5%) in all the response variables tested. Similarly, the two seed sources showed statistically significant differences throughout all the responses. The two sugarcane genotypes also showed marked variation in all the responses except the number of live buds per stalk, stalk height (cm) and a number of two bud setts produced per stalk. In sugarcane genotype B52-298, the rate of propagation for tissue culture seed source is 1:44.68 against 1:13.72 of its donor from the conventional seed source. In NCo-334, tissue culture seed source produced a propagation rate of 1:40.11 against 1:13.98 of its donor conventional seed source. Regardless of the other benefits, planting a hectare B52-298 and NCo-334 provided a direct net benefit of US$ 5448 and US$ 3999, respectively. Thus, the current result revealed that tissue culture seed source is a realistic and better alternative over the conventional seed source in sugarcane as initial seed cane at Tendaho Sugar Development Project. Evaluation of the two seed sources for all genotypes at each sugar estates/projects in the successive three-tier system of seed cane production and commercial stages could be the future line of work.