Improving beta cell isolation and purification techniques is a critical step towards the development of new cell-based therapies, diagnostic applications and diabetes research. Pancreatic Islets are composed of mixed cell populations, among them beta cells, which represent a major focus of interest due to their participation in the pathology of diabetes. Various techniques have been suggested to accomplish this step, yet efficient and robust isolation of beta cells remains a challenging task.
The present invention provides an efficient tag-free isolation method for pancreatic cell sub-types, based on separation according to a newly identified collection of surface markers. These markers are tightly correlated with specific functions, such as insulin production, ensuring enrichment of the desired functionality.
Probing against the newly identified markers in a combinatorial manner allows high degree of purity without compromising the yield, significantly increasing the amount of purified cells. Finally, the method is compatible with both extracts of pancreatic tissues and stem cells derived cultures, the latter set up high expectations in the diabetes therapy field.
A kit for isolation of distinct pancreatic cell subtypes
- High purity without compromising the yield of isolated cells.
- Compatible with a variety of heterogeneous sources including cells extracted from pancreatic tissue, committed lineages of stem cells and cultures of differentiated stem cells.
Using an innovative high throughput screen, linking specific cell surface markers with a particular functionality (e.g. insulin production), a collection of markers not previously identified in connection with pancreatic cells or with diabetes was found to be consistently expressed in human islets.
Cell isolation according to the selected markers is performed by exposing the heterogeneous source of cells to specific antibodies that recognize these markers, followed by a choice of sorting techniques such as fluorescence activated cell sorting (FACS).
The innovative concept of this method is the use of marker combinations, iterating the selection. Only cells that express both markers will be sorted out, thus increasing specificity and reducing contaminations. This increased specificity gives rise to a higher degree of purity without compromising the yield, resulting in larger amounts of isolated cells.
By applying the initial screen in yet another iteration, additional markers can be added to the selection, to refine the isolation procedure.
As this method is generally applicable to the purification of mature as well as pluripotent or partially differentiated beta cell progenitors, it holds great potential for the isolation of clinically relevant cells for treatments of diabetes.