Isolation and extraction of normal, stem and cancer cells

Isolation of Cells from Various Human and Animal Tissues

Cells can be isolated for laboratory culture using several methods. Among the most important enzymes used in the extraction process are collagenase, trypsin, dispase, and pronase, which break down the extracellular matrix. Cells that are cultured directly from the target tissue are known as primary cells (first-generation). Most cell cultures, except those derived from tumors, have a limited lifespan. An immortal cell line can be established through random or targeted mutations, allowing for indefinite proliferation and serving as a representative of a specific cell type.

Isolation of Mesenchymal Cells from Adipose Tissue

Mesenchymal stem cells (MSCs) have garnered significant attention due to their valuable properties, such as immune modulation, tissue regeneration, and multilineage differentiation potential. The application of these mesenchymal stem cells in research, particularly in clinical settings, necessitates the production of large quantities of MSCs with a low passage number within a short timeframe. Developing an optimized and standardized method for the large-scale isolation of stem cells derived from adipose tissue may provide a solution to this challenge.

This protocol presents a combined method (enzymatic digestion and mechanical disruption) as a standard, easily applicable approach to obtain significant quantities of mesenchymal stem cells. We believe that the integration of enzymatic and mechanical methods can enhance the accessible surface area of adipose tissue, thereby facilitating enzymatic digestion. Utilizing this method reduces the amount of collagenase required and has minimal impact on the viability and differentiation potential of adipose-derived mesenchymal stem cells.