To study the expression profile of miRNA122a in stem cells derived from miRNA122a positive HuH7 cell line, we maintained the cells in a stem cell enriching NSA media as 3D tumorspheres. (A) Phase contrast images of tumorspheres captured at day 5 after seeding HuH7 in NSA media. The cells were collected and total cDNA was synthesized from total cellular RNA. qRT-PCR was then performed to quantify the levels of stemness related genes (B) CD44, (C)CD133, and (D) CD90 (E) Oct4 which were reported as copy number per 10,000 copies of GAPDH house keeper using the formula (2^(Ct housekeeper- Ct sample))*10000. (F) The expression levels of miRNA122a was expressed as copy number per 10,000 copies of control. Difference between two groups was tested with a two-tailed t-test using Graph Pad Prism. (n > 3, *, p < 0.05, **, p < 0.005).