Fig. 1.

Single-cell isolation for clonal expansion. (A) Schematic representation of single-cell FACS isolation of CD133⁺CD45⁻ cells from 16-month-old Mat1a^−/− mice. On day 1, single cells were plated into individual wells of a 96-well plate. After 21 days, 10/96 wells showed colonies with growth over 50% of the well bottom. Five of these colonies were selected for expansion (CD133⁺ clone lines 1–5). (B) RT-PCR. In each clone line 1–5, coexpression of Albumin and CK-19 indicated bipotential stem cells, with strong expression of oval cell–associated transcription factors hepatocyte nuclear factor 1α and 3β (HNF1α and HNF3β) and ABCG2 transmembrane pump. Strong expression of growth factor receptors c-Met and epithelial growth factor receptor (EGFr) were also detected by RT-PCR. (C) Western blot. High levels of prominin-1 (CD133) and αFP protein were detected in each clone line (+ control = positive control for Western blot).