Methods for dynamic evolution and monitoring of characteristics in living cells using a microfluidic-enabled multi-well cell culture devices and systems

Inventors

LUDLAM, Mary J. C. • WARTMANN, David • GALLAGHER, Ciara

Assignees

Cairn Biosciences Inc

Abstract

A method for dynamic evolution and/or adaptation and monitoring of characteristics in living cells is provided, wherein the method may be performed at a microfluidic-enabled cell-culture device comprising 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. The method may involve 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, and removing all or part of the evolved/adapted population of cells from the first well.

Core Innovation

The invention provides microfluidic-enabled cell-culture devices, referred to as SmartPlate systems, with a pneumatic layer that directs flow of fluid to a plurality of individually addressable wells. One or more sensors are configured to detect data regarding environments inside one or more of the wells, enabling dynamic monitoring of conditions within a selected well. The system supports dynamic evolution and adaptation of living cell populations through repeated cycles of culturing, perturbing, monitoring, and transferring evolved material between wells.

A pneumatic vacuum/pressure routing architecture routes pneumatic actuation to microvalves and micropumps, including manifolds and manifold adapters. The plate architecture is described as modular and layered, including a well layer, a fluid routing layer, pneumatic actuation layers, sensors and a controller, along with degassers and on-chip reservoirs. This arrangement supports closed-loop operation at the per-well level by selecting and independently controlling wells while sensing one or more environmental parameters.

The method includes culturing a population of cells in a first well, perturbing one or more characteristics of an environment in the first well after culturing, and monitoring one or more characteristics of the population in the first well as it evolves. All or part of the evolved population of cells is removed from the first well and seeded into a second well separate from the first well, where a second population is cultured, perturbed, and monitored as it evolves. The document further describes examples of long-term culture and perturbation, along with sampling and recovery of evolved or adapted cells for downstream passaging or analysis.

Claims Coverage

The document includes one independent claim that covers a complete iterative workflow for dynamic evolution and monitoring of living cells across at least two individually addressable wells in a microfluidic-enabled, sensor-equipped device. The inventive features primarily define the multiwell microfluidic device structure and the repeated sequence of culturing, perturbation, monitoring, removal, seeding, and re-perturbation in a separate well.

Overall, the claim coverage centers on combining an individually addressable, pneumatic microfluidic cell-culture environment with sensors for environmental detection and a repeated evolution-and-transfer workflow between a first well and a second well, including perturbation and monitoring at each stage.

Stated Advantages

Documented Applications