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Progress in stem cell research has prioritized the refinement of cell-labeling techniques for in vitro and in vivo basic and therapeutic studies. Although quantum dots, because of their optical properties, are emerging as favorable nanoparticles for bioimaging, substantial refinements or modifications that would improve their biocompatibility are still required. We report here that internalizing quantum dots (i-QDs) generated by their conjugation with an internalizing antibody against a heat shock protein-70 family stress chaperone, mortalin, offered an efficient, genetically noninvasive, nontoxic, and functionally inert way to label mesenchymal stem cells (MSCs). The i-QD-labeled MSCs underwent normal adipocyte, osteocyte, and chondrocyte differentiation in vitro and in vivo, suggesting the potential application of i-QDs in in vivo diagnostics, regenerative and therapeutic medicine.

Adult bone marrow cells (BMCs) include two populations:;mesenchymal stem cells (MSCs), which can differentiate into bone, cartilage, and fat; and hematopoietic stem cells (HSCs), which produce all mature blood lineage. To study the effect of aging, gender, and age-related disorders on lineage differentiation, we performed quantitative RT-PCR to examine mRNA expression of the major factors defining BMC lineage, cbfa1 for osteoblasts, ppar-gamma for adipocytes, sox9 for chondrocytes, and rankl for osteoclasts, in bone marrow from 80 healthy subjects and patients (14-79 years old) with two age-related disorders: osteoarthritis (OA) and rheumatoid arthritis (RA). Two apoptosis-related genes, bcl-2 and drak1, were studied. RANKL and PPAR-Gamma levels exhibited a clear positive correlation with age in female patients, but not in males, with a slight age-related decline in CBFa1 transcripts. DRAK1 expression showed an age-associated ascending trend with significantly greater transcripts of RANKL and DRAK1 in females (p < 0.01). Compared with age-matched controls, RA patients exhibited increased RANKL, PPAR-Gamma, and DRAK1 mRNA levels (p < 0.05), and OA showed the higher RANKL and PPAR-Gamma transcripts (p < 0.05). Furthermore, SOX9 and DRAK1 expressions in the RA group were higher than in the OA group (p < 0.05). Our data indicate that aging and age-related disorders affect gene expressions differently, suggesting that in aging, the lineage of bone marrow cells was modified with prominent changes in decreased bone marrow osteoblastogenesis, increased adipogenesis and osteoclastogenesis, while in age-related disorders, marrow adipogenesis and the activity or number of osteoclasts may play an important role in the pathogenesis of arthritic bone loss.

Nanopillar sheets were fabricated with high-aspect ratio structures with a diameter of 160-1,000 nm and a height of 1 mum by nanoimprinting. The suitability of nanopillar sheets as a new type of cell culture dish was examined by studying the behavior of HeLa cells cultured on the sheets using light microscopy, scanning electron microscopy, and fluorescence microscopy observing actin and vinculin molecules. The nanopillar structure enabled easy subculture of the cells from the sheets without conventional trypsinization. Moreover, the HeLa cells divided and proliferated on the sheets in a different way to that found on petri dish because of the manner in which the cells adhered to the materials.

This is the first report of the successful culturing of HeLa cells on nanopillar sheets–a new type of cell culture dish–in a different way from that on flat petri dishes. Nanopillar sheets were fabricated with a high-aspect ratio structure with a diameter of 80–1000 nm and a height of 1–3 µm using nanoprint technology. Nanopillar structure with 500 nm diameter and 1 µm height enabled easy subculture of the cells from the sheets without the conventional trypsinization method. Moreover, the HeLa cells divided and proliferated on the sheets in the same way as on the flat surfaces with different manner of adhesion.