Mesenchymal stem cells (MSCs) are known to have the capacity for self-renewal and differentiation into mesenchyme-lineage cell types, including osteoblasts, adipocytes, chondrocytes, tenocytes, and myoblasts, and contribute to the regeneration of a variety of mesenchymal tissues. Adipose tissue represents a rich source of mesenchymal stem cells, and provides an abundant and accessible source of adult stem cells with minimal patient discomfort. Isolation of adipose tissue-derived stem cells (ADSCs) has primarily been achieved with Liberase HI, which is composed of Clostridium histolyticum collagenases I and II and thermolysin. All Liberase preparations contain endotoxin, which is harmful for cell viability. The recombinant production of Liberase enzymes is marred by the instability of collagenase I. The matrix metalloproteinases (MMPs) are a family of enzymes capable of catalyzing the degradation of virtually all ECM components, including collagens, laminin, fibronectin, and elastin. There are no literature studies describing the use of MMPs for the isolation of stem cells from adipose tissue. The research plan described herein focuses on the exploration of recombinant MMP cocktails for the efficient isolation of ADSCs. To achieve this goal the specific aims are (1) evaluate the efficacy of MMP-induced isolation of stem cells from adipose tissue and (2) evaluate MMP versus bacterial collagenase efficiencies for stem cell isolation. The applied research and production of recombinant MMPs proposed in this application is very significant, because without the highest quality of connective tissue degrading enzymes it is virtually impossible to liberate viable ADSCs with good and stable proliferative capabilities. The overall impact of the proposed research and applications in the field of stem cell transplantation and advancements in treatment of a broad array of diseases is expected to be significant.
The present study is designed to create a new method for the isolation of stem cells from adipose tissue. The overall impact of the proposed research and applications in the field of stem cell transplantation and advancements in treatment of a broad array of diseases is expected to be significant.