Case Study

Cloning Of Three Cell Types Comparing The Namo™ Single Cell Dispenser To Flow Cytometry And Limited Dilution Cloning

Michael Brem1, Thi Lam1, April Stanley1, John Mott1, Jesse McCool1, Soohee Cho2, Abe Couse2, Junyu Lin2
1 Cytovance Biologics, 800 Research Parkway, Oklahoma City, OK 73104
2 Namocell, Inc, 2483 Old Middlefield Way, Mountainview, CA 94043

The FDA requires commercial biotherapeutic manufacturing to use an expression cell line derived from a single cell or clone [1]. Limiting dilution cloning (LDC) has been the standard method established for generating clonal cell lines, based on statistical data [2,3], however it is time-consuming and can be inefficient. Recent advances in imaging technology have provided the ability to directly visualize proof of clonality, however LDC, when used with imaging technology, can result in too few or too many cells in each of the wells. A Fluorescence-activated Cell Sorter (FACS) can dispense a single cell in each well of a plate [3], but high fluidics pressure can reduce cell viability and FACS devices are very costly. The Namo™ Single Cell Dispenser was designed by Namocell® to circumvent some of these disadvantages by seeding a single cell into each well of a plate, using much lower fluidics pressure [4], resulting in a higher number of wells containing viable single cells. Here we compared each of these methods, using different cell lines, to determine which method would deliver the highest number of viable clones. The results of our study show the Namo™ Single Cell Dispenser delivered the highest number of wells containing viable single cells when compared to LDC or FACS for all cell lines included.

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