Cell-chip and automatic controlled system capable of detecting conditions for optimizing differentiation of stem cell using mechanical stimulus

• US 8,187,863 B2
• Filed: 09/24/2008
• Issued: 05/29/2012
• Est. Priority Date: 02/29/2008
• Status: Active Grant

• Current Assignee: Ajou University Industry
• Original Assignee: Ajou University Industry
• Inventors: Sim, Woo Young, Yang, Sang Sik
• Primary Examiner(s): Beisner, William H
• Assistant Examiner(s): HOBBS, MICHAEL L
• Application Number:
  US12/438,103
• Publication Number:
  US 20100233791A1
• Time in Patent Office: 1,343 Days
• Field of Search: 435/286.4, 435/287.1, 422/72
• US Class Current: 435/286.4
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/146 Employing pressure sensors
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/0803 Disc shape
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0874 Three dimensional network
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/14 Means for pressure control
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/0655 pinch valves
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: C12M 23/12 Well or multiwell plates C12M25/04 takes precedence
• CPC Class Codes: C12M 23/16 Microfluidic devices; Capillary tubes integrated microfluidic structures B01L3/5027; microreactors B01J19/0093
• CPC Class Codes: C12M 35/04 Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli
• CPC Class Codes: G01N 2035/00158 Elements containing microarrays, i.e. "biochip"
• CPC Class Codes: G01N 2035/1044 Using pneumatic means
• CPC Class Codes: G01N 35/1074 arranged in a two-dimensional array
• CPC Class Codes: G01N 35/1095 for supplying the samples to flow-through analysers for a specific analyser see relevant groups, e.g. under G01N15/00, G01N21/00, G01N27/00, G01N30/00, H01J49/00

Abstract

Provided are a cell chip and a system thereof that are capable of detecting optimal conditions for stem cell differentiation by mechanical stimuli. The cell chip for cell differentiation experimentation includes a plurality of cell chambers for storing cells and culture media, cell and culture medium injection ports for transferring the cells and culture media to corresponding cell chambers, fine passages for moving the cells and the culture media injected into the cell and culture medium injection ports to the cell chambers, pneumatic injection ports for injecting pneumatic pressures applied to the cell chambers, and apertures having circular films for transferring the pneumatic pressures injected through the pneumatic injection ports to corresponding cell chambers. Here, at least two of the apertures may have different areas to vary the magnitude of pneumatic pressure applied to corresponding cell chambers.

Claims

1. A cell chip for cell differentiation experimentation of stem cell, comprising:

a plurality of cell chambers for storing cells and culture media;

cell and culture medium injection ports for transferring the cells and culture media to corresponding cell chambers;

first fine passages for moving the cells and the culture media injected into the cell and culture medium injection ports to the cell chambers;

a pneumatic injection port for injecting pneumatic pressures applied to the cell chambers;

apertures having circular films for transferring the pneumatic pressures injected through the pneumatic injection ports to corresponding cell chambers;

marker and dye injection ports for injecting a marker or dye to observe a cell state and a differentiation process in the cell chamber;

second fine passages connected between the marker and dye injection ports and acting as a moving passage of a marker or dye to the cell chamber; and

a micro-valve device for adjusting fluid flow between the fine passage and the cell chamber, fluid flow between the passage and the cell chamber, and fluid flow between two adjacent cell chamber,

wherein;

at least two of the apertures have different areas to vary the magnitude of pneumatic pressure applied to corresponding cell chambers;

the plurality of cell chambers are disposed on a plurality of concentric circles about the pneumatic injection port, at least two cell chambers being disposed on a same circle about a center of the pneumatic injection port, the plurality of cell chambers are divided into a plurality of units, each unit consisting of a number of cell chambers, between two adjacent ones of which the first fine passages are disposed; and

the micro-valve device comprises;

first micro-valve pneumatic injection ports for injecting first micro-valve pneumatic pressure used to adjust flow between the cell chamber and the first fine passage and to adjust flow between two adjacent ones of the cell chambers in each unit;

second micro-valve pneumatic injection ports for injecting second micro-valve pneumatic pressure used to adjust flow of a marker or dye between the cell chamber and the second fine passage;

first pneumatic chambers disposed to be in contact with the first fine passage between corresponding cell chambers and contracting and expanding in response to the first micro-valve pneumatic pressure;

second pneumatic chambers disposed to be in contact with each corresponding second fine passage for a marker or dye and contracting and expanding in response to the second micro-valve pneumatic pressure;

a first valve boss disposed between each corresponding cell chamber and first fine passage and connected to a corresponding first pneumatic chamber to be opened or closed in response to contraction or expansion of the corresponding first pneumatic chamber, wherein flow between two adjacent ones of the cell chambers in each unit is blocked when the first valve boss is closed; and

a second valve boss disposed between each corresponding cell chamber and second fine passage for a marker or dye and connected to a corresponding second pneumatic chamber to be opened or closed in response to contraction or expansion of the corresponding second pneumatic chamber.

2. The cell chip according to claim 1, further comprising:

a pressure sensor disposed at a lower surface of the cell chamber and sensing pressures of the cell chambers,

wherein the lower surface of the cell chamber is a surface opposing a surface disposed in a direction from which pneumatic pressure is applied to the cell chambers.

3. The cell chip according to claim 2, wherein the pressure sensor comprises:

first electrodes formed of indium tin oxide (ITO), disposed in the same arrangement as the cell chambers, and mounted on lower surfaces of the corresponding cell chambers;

second electrodes formed of ITO and disposed opposite to the corresponding first electrodes; and

gap disposed between the first electrodes and second electrodes.

4. The cell chip according to claim 1,

wherein the apertures have different areas according to distance from the pneumatic injection port of the corresponding cell chamber.

5. A cell chip control system, on which a cell chip according to claim 1 is mounted, for supplying pneumatic pressure, a cell, or a culture medium to the cell chip, and observing and measuring a state and differentiation of the cell in the cell chip.

6. The cell chip control system according to claim 5, comprising:

an X-Y stage having a mounting part on which a cell chip is mounted, and moving or rotating the mounting part in a first direction in response to a first position signal;

a Z stage moving in a second direction depending on a second position signal and connecting connection pipes for supplying pneumatic pressure, a cell, and a culture medium to the cell chip to an injection port corresponding to the cell chip;

a pneumatic valve for varying magnitude and supply duration of pneumatic pressure supplied to a corresponding connection pipe in response to a first control signal;

a syringe pump for providing a cell and a culture medium to the corresponding connection pipe in response to a second control signal;

a measurement circuit for supplying a cell, a culture medium, and pneumatic pressure to the cell chip and measuring pressures of the cell chambers of the cell chip after an arbitrary time elapses; and

a microcomputer for generating the first position signal, the second position signal, and the control signal, and receiving a measurement value from the measurement circuit to transfer the value to a user through an input/output device.

7. The cell chip control system according to claim 6, further comprising:

a fluorescence microscope disposed at an upper part or a lower part of the mounting part and enabling visual observation of a cell in the cell chamber of the cell chip using a marker or dye.