US 12,258,551 B2
Methods for dynamic evolution and monitoring of characteristics in living cells using a microfluidic-enabled multi-well cell culture devices and systems
Mary J. C. Ludlam, San Francisco, CA (US); David Wartmann, Berkeley, CA (US); and Ciara Gallagher, Redwood City, CA (US)
Assigned to Cairn Biosciences, Inc., San Francisco, CA (US)
Appl. No. 17/293,407
Filed by Cairn Biosciences, Inc., San Francisco, CA (US)
PCT Filed Nov. 14, 2019, PCT No. PCT/US2019/061514
§ 371(c)(1), (2) Date May 12, 2021,
PCT Pub. No. WO2020/102562, PCT Pub. Date May 22, 2020.
Claims priority of provisional application 62/768,493, filed on Nov. 16, 2018.
Prior Publication US 2021/0403853 A1, Dec. 30, 2021
Int. Cl. C12M 1/34 (2006.01); C12M 1/00 (2006.01); C12M 1/42 (2006.01); C12M 3/06 (2006.01); C12N 5/09 (2010.01); G01N 21/64 (2006.01)
CPC C12M 41/36 (2013.01) [C12M 23/16 (2013.01); C12M 23/20 (2013.01); C12M 29/00 (2013.01); C12M 35/04 (2013.01); C12M 35/08 (2013.01); C12M 41/46 (2013.01); C12M 47/04 (2013.01); C12N 5/0693 (2013.01); G01N 21/6428 (2013.01); G01N 21/6452 (2013.01); C12N 2500/00 (2013.01); C12N 2527/00 (2013.01); G01N 2021/6439 (2013.01)] 38 Claims
 
1. A method for dynamic evolution and monitoring of characteristics in living cells, comprising:
in a microfluidic-enabled cell-culture device comprising a pneumatic layer for directing flow of fluid to a plurality of individually addressable wells, and one or more sensors configured to detect data regarding environments inside one or more of the plurality of wells:
culturing a population of cells in a first well of the plurality of wells;
perturbing one or more characteristics of an environment in the first well following the culturing of the population of cells;
monitoring one or more characteristics of the population of cells in the first well;
removing all or part of the evolved population of cells from the first well;
seeding a second well, separate from the first well, with the all or part of the evolved population of cells;
culturing a second population of cells, comprising all or part of the evolved population cells, in the second well;
perturbing one or more characteristics of an environment in the second well following the culturing of the second population of cells; and
monitoring one or more characteristics of the second population of cells as it evolves in the second well.