The potential of stem cells to replace damaged or malfunctioning tissues has been studied for some time, and recent studies using adipose‐derived mesenchymal stem cells (ADSCs) have shown promise for adult stem cells as a rich, clinically‐relevant population. Our lab has previously determined that the quantity and quality of ADSCs varies depending on patient age and sex, anatomical location of harvest and whether it was a previously treated site with lipoplasty. A clinically‐relevant question is whether method of lipoplasty affects pluripotency of the stem cells. Our lab has shown that harvesting ADSCs using BodyJet and BodyJet with SmartLipo lipoplasty techniques produces a high yield of viable stem cells. Using established plating densities, corrected for stem cell viability measured by flow cytometry, we have reared isolated ADSCs, confirmed by flow cytometry and confocal microscopy, harvested with the BodyJet and BodyJet with SmartLipo techniques. This result suggests that both BodyJet and BodyJet with SmartLipo lipoplasty techniques yield clinically useful ADSC's. Funded, in part, under a grant with the Pennsylvania Department of Health and intramural funds.

Clinical use of stem cells provide undeniable advantages as they can substitute for injured cells, tissues, and even organs in the human body. Adult adipose‐derived stem cells (ADSCs), have vast potential for clinical medicine because they can easily be harvested from a patient and provide an autologous transplant option. Data gathered in our lab has shown that harvesting ADSCs using water‐jet assisted liposuction (BodyJet) with laser‐assisted lipoplasty (SmartLipo) techniques produces a high yield of viable stem cells. These isolated cells can be plated in monolayer culture and retain their stem cell phenotype. In this study, we test the differentiation abilities of these BodyJet/SmartLipo harvested cells following cryopreservation. Since the clinical use of ADSCs is highly indicated for orthopedic and arthritis applications, we tested the ability of the liposuction‐harvested ADSCs to differentiate along the chondrocyte and osteoblast lineages. In this study, we compare the cryopreserved primary human ADSCs to a normal human, mesenchymal ADSC line, to measure stem cell viability and differentiation following thaw. Utilizing confocal microscopy, real‐time PCR, cell counts, alizarin red and alcian blue staining, the stem cells show promise in their ability to differentiate along the chondrocyte and osteoblast lineages. These data support the clinically‐relevant idea that lipoplasty‐obtained ADSCs will retain multipotency. Grant Funding Source: Supported by Pennsylvania Department of Health

Our previous work demonstrated the sensitivity of osteoblasts to the presence of microenvironmental apoptogens. As osteoblasts change their functional orientation with maturation, it follows that the sensitivity of these cells to apoptosis must change as well. It is the goal of this study to exam the relationship between the maturation of osteoblasts and the susceptibility of these cells to apoptosis. For this study the Human Fetal Osteoblast (hFOB) cell line was used. These cells exhibit an early osteoblastic phenotype when cultured at 34ºC while above 37ºC, the cells mature. Cells were cultured for 2, 7 and 14 days. Phenotypic changes were confirmed. Cells cultured at both 34ºC and 39.5ºC were exposed to increasing levels of the calcium and phosphate ion pair. Percentages of viable cells were measured using the MTT assay. Apoptosis was confirmed by evaluating Caspase‐3 activation and TUNEL staining. Results indicated that hFOB cells incubated at 39.5ºC exhibited a greater sensitivity to the activation of apoptosis by the calcium and phosphate ion pair. These results suggest that with maturation, osteoblasts become more sensitive to the activation of apoptosis. This work was supported by a grant from the Center for Chronic Disorders of Aging at the Philadelphia College of Osteopathic Medicine.

Our previous work demonstrated the sensitivity of bone cells to microenvironmental apoptogens. As the lifespan of an osteocyte can extend into decades, osteocytic terminal differentiation must necessarily remove such sensitivity. Therefore, the goal of this study is to probe the relationship between osteocyte differentiation state and sensitivity to apoptosis. Murine Long bone‐derived osteocytes (MLO) A5 and Y4 cell lines, representative of early and late stage osteocytes, respectively, were seeded onto well plates, allowed to grow for 2, 7 or 14 days and subsequently exposed to increased levels of the calcium and phosphate ion pair for 24 h. Following exposure, MTT assays were performed to quantify cell death. Apoptosis was confirmed through caspase‐3 and TUNEL staining. Osteocytic phenotypes were confirmed through alkaline phosphatase staining. Results indicated that, when treated with increased levels of the calcium and phosphate ion pair, the MLO A5 cell line, representative of early osteocytes, exhibited greater sensitivity to apoptosis than the MLO Y4 cell line, representative of late osteocytes. These data suggest that sensitivity to calcium and phosphate induced apoptosis is a function of maturation level. This work was supported by a grant from the Center for Chronic Disorders of Aging at the Philadelphia College of Osteopathic Medicine.