The isolation of primary canine satellite cells utilizing microarray technologies

The isolation of specific cell populations from a heterogeneous cell sample is of great interest to the biomedical community, particularly in the areas of cell-based therapeutics. These therapeutic techniques rely upon the use of pure cell populations for the treatment and possible cure of a number of potentially lethal diseases. Current biological technologies present limited utility for cell isolations, particularly when cellular identification methods require destructive assays. To this end, it is imperative to provide techniques and technologies capable of identifying, isolating, and collecting desired cells for future use.;Two microarray technologies, referred to as micropallets and microrafts, have recently been introduced for the isolation and identification of adherent cells. Micropallet arrays are micron-scale pedestals fabricated on glass substrates, while microrafts are of similar design yet are held within an elastomeric polymer microwell array. Following the culture of adherent cells on the microarray elements, individual elements possessing the cultured cells can be detached and isolated from the array. These technologies have demonstrated the successful sorting of cells from heterogeneous cell samples with high viability post-sorting. However, previous microarray technologies lacked the ability to culture primary cells, limiting their utility to the biological community.;In this dissertation, microarray technologies are described that enabled the culture, isolation, and identification of cells. First, the modification of micropallets is described for the culture of primary cells. The addition of polymer coatings to micropallet surfaces permitted the culture and sorting of primary canine satellite cells with no adverse effects to cell physiology. Microwell arrays were then developed to enable the selective collection of released micropallets using a laser-based release method. Through the detailed analysis and optimization of micropallet release trajectories and travel distances, an efficient collection system was developed enabling the isolation of hundreds of micropallets into desired microwells and was demonstrated for cell sorting with high yield, purity, and cell viability. Finally, a two-part micromolded array-based technology utilizing microrafts was demonstrated for the culture, sampling, identification, and expansion of primary canine satellite cells.