Behavior-activated cell selection: Application to stem cell expansion

BIOT 1

Brad Dykstra1, John Ramunas2, David Kent1, Connie J. Eaves1, and Eric Jervis, ericjj@cape.uwaterloo.ca2. (1) Terry Fox Laboratory, British Columbia Cancer Agency, 675 West 10th Avenue, Vancouver, BC V5Z 1L3, Canada, (2) Department of Chemical Engineering, University of Waterloo, 200 University Ave. West, Waterloo, ON N2L 3G1, Canada
The nondestructive nature of cellular behavior analysis using long-term imaging facilitates cell classification. To look for new indicators of self-renewing capacity, hematopoietic stem cells (HSCs) isolated from adult mouse marrow were individually micromanipulated into the wells of a microarray imaging chamber and then cultured for 4 days with each cell and its progeny being imaged using time-lapse photography. Individual clones were then harvested and assayed in mice for HSC compartment repopulating activity. Characteristics identified from image sequences were screened and 3 were found to be predictive of HSC potential: mitotic activity, longer cell cycle times, and uropodia. Clone selection criteria developed using these data allowed all HSC-containing clones to be detected and at a 2.4-fold greater efficiency in subsequent experiments. The BACS platform represents an advance towards the goal of dynamic single cell analysis in heterogeneous populations and demonstrates the potential of this technology for preparative cell isolation devices.