Title

Adipose Derived Stem Cells (ADSC) Harvested from Lipoaspirate Differentiate into Chondrocytes and Osteoblasts

Date of Award

7-2014

Degree Type

Thesis

Degree Name

Master of Science (MS)

First Advisor

Marina D'Angelo, PhD

Second Advisor

Christopher Adams, PhD

Third Advisor

Ruth Borghaei, PhD

Fourth Advisor

Marcus G Bell, PhD

Abstract

Clinical use of stem cells provides 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 (Body Jet) with laser assisted lipoplasty (SmartLipo) techniques produces a higher yield of viable stem cells compared to other commonly-used lipoplasty techniques. These isolated cells can be plated in monolayer culture to retain their stem cell phenotype. In this study, we test the ability of cells derived from cryostored lipoaspirate to differentiate along the chondrocyte and osteoblast lineages. Cells isolated from cryopreserved lipoaspirate collected by Body Jet or Body Jet in combination with SmartLipo were compared to a human, mesenchymal ADSC line (PCS-500-011). Differentiation parameters were optimized in the ADSC cell line including measurement of alkaline phosphatase activity, alizarin red and alcian blue staining of matrix mineralization, and qRT-PCR or immunocytochemistry to measure matrix-specific molecule production. The ADSC line differentiated into osteoblasts and chondrocytes by 2 weeks in culture, but the chondrocyte lineage began to de-differentiate between weeks 2 and 4 in culture. Utilizing confocal microscopy, we demonstrated that following isolation from frozen lipoaspirate, cells reared in chondrocyte differentiation media or osteoblast differentiation media could differentiate along the chondrocyte and osteoblast lineages. These data support the clinically relevant notion that stem cells retrieved from cryostored lipoaspirate will retain pluripotency.

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