The basic factors affecting the success of tissue culture are the origin and size of the explants. Therefore this work was conducted at commercial tissue culture Laboratory), Belbis City, Sharkia Governorate, Egypt since 2016 till 2018, to evaluate two different explants sources Local Explants (LE) and Imported Germaplasm(IG) and three explants sizes (small, medium, large explants) of bananas cv Grand Nine through tissue culture technique. Four stages were carried out (starting, multiplication, rooting, and acclimatization stages). The obtained results indicated that LE with large sizes had higher and significant number of shoots length and diameter at the end of multiplication stage in comparison with IG; also LE with small or medium sizes had higher rooting characteristics than IG, moreover at the end of acclimatization stage LE with large size of explant had significant length, girth, and number of leaves per plant, with low number of mutant plants in comparison with IG. Therefore LE could be used for banana Grand Nine cv micropropagation with higher growth and rooting characteristics with low number of mutant plants and with low coast of preparation instead of IG.