Microfluidic Cell Culture With Segmented Sterile Access Ports

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Solution Overview

Problem

Current methods for culturing cells in microfluidic devices face challenges in providing a suitable environment for cell growth, viability, and portability, as they lack efficient techniques for precise cell handling, environmental control, and isolation.

Innovation Solution

A microfluidic device with a flow region and growth chambers, where the growth chambers have surfaces conditioned with specific moieties such as polymers and cell adhesion blocking molecules, and include a dielectrophoresis configuration for cell manipulation, allowing for controlled diffusion of media and optimal cell growth conditions.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If conventional cell culture methods are used, then cell growth and proliferation can be achieved, but contamination by bacteria and mycoplasma occurs and experimental reproducibility is compromised

Engineering Contradiction:
Improveexperimental reproducibilityVSAvoidcontamination
Core Design Contradiction:
ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The invention divides the cell culture system into isolated micro-wells within a single well plate, creating physically separated culture environments. Each micro-well acts as an independent containment unit with its own barrier layer, preventing cross-contamination between samples while maintaining sterile conditions through integrated access ports that eliminate repeated opening/closing operations.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The invention introduces an access port system as an intermediary mechanism that allows media exchange and cell manipulation without breaking sterile barriers. The access ports with caps enable controlled access to individual micro-wells while maintaining the integrity of the barrier layers, serving as a mediator between the external environment and the sterile culture environment.

Inventive Principle:
Principle #24Intermediary (Mediator)

2Adaptability or versatility

If multiple cell lines are cultured separately in conventional flasks, then each cell line can be maintained, but the device complexity and space requirements increase

Engineering Contradiction:
Improvemulti-cell line capabilityVSAvoidnumber of separate containers
Core Design Contradiction:
Adaptability or versatilityVSDevice complexity

Solution Approach 1:

The invention merges multiple separate cell culture operations into a single integrated well plate containing multiple micro-wells. Each micro-well functions as an independent culture chamber, allowing simultaneous cultivation of different cell lines or conditions in one device, thereby reducing the total number of separate containers and simplifying the overall system architecture.

Inventive Principle:
Principle #5Merging (Combining)

Solution Approach 2:

The invention transitions from horizontal arrangement of separate flasks to a two-dimensional grid structure within a single well plate. By organizing multiple micro-wells in rows and columns, the system efficiently utilizes space in a compact format, enabling high-throughput culturing without increasing vertical space requirements or device complexity.

Inventive Principle:
Principle #17Another dimension (Dimensionality change)

3Reliability

If frequent media changes are performed in conventional culture, then cell health is maintained, but the risk of contamination increases with each opening

Engineering Contradiction:
Improvesterility maintenanceVSAvoidfrequency of access operations
Core Design Contradiction:
ReliabilityVSLoss of time

Solution Approach 1:

The invention implements preliminary sterilization and sealing of the entire well plate system before cell culture begins. The barrier layers are pre-assembled and sealed, creating a sterile environment that is maintained throughout the experiment. Media and reagents are prepared and sterilized in advance, eliminating the need to break sterile barriers during routine media changes.

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

The access ports with removable caps serve as intermediaries that enable media exchange without compromising sterility. The cap acts as a temporary barrier that can be opened for media addition and then resealed, maintaining the sterile environment while allowing necessary interventions. This intermediary mechanism decouples the frequency of media changes from the risk of contamination.

Inventive Principle:
Principle #24Intermediary (Mediator)

Data Source

PatentEP3286296A1Microfluidic cell culture
Publication Date: 2018.02.28 BERKELEY LIGHTS INC

AI summary

Systems, methods and kits are described for culturing one or more biological cells in a microfluidic device, including provision of nutrients and gaseous components configured to enhance cell growth, viability, portability, or any combination thereof. In some embodiments, culturing a single cell may produce a clonal population in the microfluidic device.