PT  - JOURNAL ARTICLE
AU  - Teranishi, Kazuki
AU  - Wagatsuma, Keisuke
AU  - Toda, Keisuke
AU  - Nomaru, Hiroko
AU  - Yanagihashi, Yuichi
AU  - Ochiai, Hiroshi
AU  - Akai, Satoru
AU  - Mochizuki, Emi
AU  - Onda, Yuuki
AU  - Nakagawa, Keiji
AU  - Sugimoto, Keiki
AU  - Takahashi, Shinya
AU  - Yamaguchi, Hideto
AU  - Ota, Sadao
TI  - Label-free ghost cytometry for manufacturing of cell therapy products
AID  - 10.1101/2023.07.09.545884
DP  - 2023 Jan 01
TA  - bioRxiv
PG  - 2023.07.09.545884
4099  - http://biorxiv.org/content/early/2023/07/21/2023.07.09.545884.short
4100  - http://biorxiv.org/content/early/2023/07/21/2023.07.09.545884.full
AB  - Automation and quality control (QC) are critical in manufacturing safe and effective cell and gene therapy products. However, current QC methods, reliant on molecular staining, pose difficulty in in-line testing and can increase manufacturing costs. Here we demonstrate the potential of using label-free ghost cytometry (LF-GC), a machine learning-driven, multidimensional, high-content, and high-throughput flow cytometry approach, in various stages of the cell therapy manufacturing processes. LF-GC accurately quantified cell count and viability of human peripheral blood mononuclear cells (PBMCs) and identified non-apoptotic live cells and early apoptotic/dead cells in PBMCs, T cells and non-T cells in white blood cells (WBCs), activated T cells and quiescent T cells in PBMCs, and particulate impurities in PBMCs. The data support that LF-GC is a non-destructive label-free cell analytical method that can be used to monitor cell numbers, assess viability, identify specific cell subsets or phenotypic states, and remove impurities during cell therapy manufacturing. Thus, LF-GC holds the potential to enable full automation in the manufacturing of cell therapy products with reduced cost and increased efficiency.Competing Interest StatementS.O. is the founder and shareholder of ThinkCyte Inc., a company engaged in the development of an ultrafast imaging cell sorter. K.T., K.T., K.W., H.N., Y.Y., S.A., E.M., Y.O. and K.N. have shares of stock options from ThinkCyte Inc. K.T., K.T., K.W., H.N., Y.Y., S.A., E.M., Y.O. and K.N. are employees; H.O. and K.S. are former employees; and S.O. is a board member of ThinkCyte Inc. K.T., K.T., K.N., K.S. and S.O. filed patent applications related to the LF-GC method. S.T. and H.Y. are employees of Astellas Pharma, Inc.