<p>The continuous battle of yam (<i>Dioscorea rotundata</i> Poir) with pests and diseases as well as other environmental factors has made the crop expensive to produce. Tissue culture (<i>in vitro</i>) offers a substitute for propagating the crop, although more farmers prefer landraces of yam, which perform poorly in tissue culture. To establish an efficient and genotype-independent prolific tissue culture system in yam varieties, several hormonal supplementations in Murashige and Skoog’s (MS) complete medium were investigated on the <i>in vitro</i> growth of an improved variety and two landraces of Guinea yam. The first objective was tailored to study the effect of hormone-free MS complete medium versus optimized MS medium (supplemented with 0.1&#xa0;mg L⁻¹ of kinetin), while the second objective investigated different levels of 6-benzylaminopurine (BAP) in MS medium on the <i>in vitro</i> growth of Guinea yam. The experimental design for the first objective was simple completely randomized (CRD) with three replications. For the second objective, the experimental design was CRD arranged in 3 × 6 factorial treatment structure with three replications. Significant differences (<i>p</i> &lt; 0.05) were observed in the growth effects between the various varieties as well the different concentrations of MS medium supplemented with BAP. Dente and Pona (landraces) both performed slightly better on the hormone-free MS medium than they did on the optimized media. On the other hand, the improved variety Kukrupa performed slightly better on the optimized medium than it did on hormone-free MS medium. MS medium supplemented with different concentrations of BAP had a poor effect on the growth performance of all three varieties. The present study has revealed that hormone-free MS medium should be used in the <i>in vitro</i> propagation of Dente and Pona yams, while optimized MS medium supplemented with plant growth regulators should be used in the <i>in vitro</i> propagation of Kukrupa improved yam variety.</p>