FIG. 1.

Functional characterization of mesenchymal stromal cells (MSCs) from bone marrow (BM), normal cervix (NCx), and cervical cancer (CeCa). MSCs from the three sources (BM, n=5; NCx, n=5; CeCa, n=5) were cultured in the adipogenic, osteogenic, and chondrogenic induction medium for 14, 21, and 28 days, respectively. Adipogenic differentiation was indicated by accumulation of neutral lipid vacuoles that stained with Oil Red O. Arrows indicate cytoplasmic granules positive to staining. Scale bar=20 μm, (A−C). Osteogenic differentiation was indicated by calcium deposition, which stained with von Kossa dye. Scale bar=20 μm, (D−F). Chondrogenic differentiation was indicated by the chondrogenic matrix colored by Alcian blue in cryosections from pelleted micromass. Scale bar=200 μm, (G−I). Chondrogenic differentiation was analyzed by morphology and collagen content in cryosections from pelleted micromass. Scale bar=200 μm and 20 μm, (J−L) and (M−O panels are inset from J−L), respectively. In samples from BM, the presence of small clusters of cells, surrounded by a condensed matrix located in the same area, is evident [hyaline cartilage; (J) and (M, inset from J)]. In samples from NCx and CeCa, numerous collagenous fibers, colored by Mason's trichromic dye (blue), are visible as large irregular bundles between groups of chondrocytes [fibrocartilage; (K), (L), (N, inset from K), and (O, inset from L)]. One representative experiment is showed. In (P, Q), detection and typing of HPV were performed by PCR assay. Amplification pattern shows a conserved region of 450 bp in the L1 HPV open reading frame obtained after amplification on CeCa tissue. No bands were detected in BM-, NCx-, and CeCa-derived MSCs (BM, n=5; NCx, n=5; CeCa, n=5) (P). The CaSki cell line positive for HPV16⁺ infection was used as a positive control and the Hela cell line HPV18⁺ was used as a negative control. β-actin (230 bp) was used as an internal control (Q). One representative experiment is showed.