Systems, methods, and apparatus for induced pluripotent stem cell isolation and combinatorial production

Abstract

Described herein are various systems, methods, and apparatus for systematic creation of isolated homogeneous colonies of cells from vector-based lineages. The vector-based lineages may originate from multiple types of viral vector families (e.g., Paramyx-oviridae, Retroviridae, Parvoviridae) or non-natural engineered vectors or a plurality of vector combinations, for example. In certain embodiments, the isolated homogeneous colonies of cells are vector-free sub-colonies; in other embodiments, the isolated homogeneous colonies of cells are homogeneous vector sub-colonies. In other embodiments, vector mixed sub-colonies are created. The disclosed systems, methods, and apparatus are useful for inducible pluripotent stem cell (iPSC) production and work by selectively binding to one or more corresponding protein markers expressed on the surface of a cell that indicate that cellular reprogramming has occurred. Software is used to automate the purification and isolation of the iPSCs produced.

1 .- 13 . (canceled) 
     
     
         14 . A method for electronically determining and compiling a list of binding agents from a storage facility comprising one or more electronic files, the method comprising:
 receiving, by a processor of a computer device, data from the detection of a plurality of cell surface markers expressed on the surface of iPSCs;   determining, by the processor, one or more vectors contacted to the colony of cells based on the detected cell surface markers;   mapping, by the processor, the determined one or more vectors to a list of a plurality of cell surface markers contained in the one or more files of the storage facility; and   mapping one or more binding agents contained in an electronic compendium having a binding specificity for each mapped cell surface marker.   
     
     
         15 . (canceled) 
     
     
         16 . A method comprising:
 providing a heterogeneous mixture of a plurality of colonies of cells having been contacted with a plurality of vectors;   producing a colony of iPSCs from the plurality of colonies of cells, wherein the colony of iPSCs comprises varied genetic edits; and   separating the colony of iPSCs into one or more homogeneous vector lineages.   
     
     
         17 . (canceled) 
     
     
         18 . (canceled) 
     
     
         19 . The method of  claim 16 , wherein separating the iPSCs into one or more homogeneous vector lineages comprises:
 separating the iPSCs into cell surface marker-based lineages determined using one or more members selected from the group consisting of single bead separation, column chromatography, serial separation, microfluidic channel separation, magnetic bead binding, fluorescence activated cell (FACS) sorting, and antibody binding.   
     
     
         20 .- 30 . (canceled)