EP1244770A2 - Procede pour transferer des molecules dans des cellules - Google Patents

Procede pour transferer des molecules dans des cellules

Info

Publication number
EP1244770A2
EP1244770A2 EP00990583A EP00990583A EP1244770A2 EP 1244770 A2 EP1244770 A2 EP 1244770A2 EP 00990583 A EP00990583 A EP 00990583A EP 00990583 A EP00990583 A EP 00990583A EP 1244770 A2 EP1244770 A2 EP 1244770A2
Authority
EP
European Patent Office
Prior art keywords
source
molecules
acoustic pulses
medium
cells
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP00990583A
Other languages
German (de)
English (en)
Inventor
Friedrich Ueberle
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Dornier Medizintechnik GmbH
Original Assignee
Dornier Medizintechnik GmbH
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from DE1999162904 external-priority patent/DE19962904A1/de
Application filed by Dornier Medizintechnik GmbH filed Critical Dornier Medizintechnik GmbH
Publication of EP1244770A2 publication Critical patent/EP1244770A2/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
    • C12M35/04—Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli

Definitions

  • the invention relates to a device according to the preamble of claim 1
  • Sonotrodes must first be brought into resonance in order to generate vibrations which have such a high amplitude that exceeds the cavitation threshold. In a disadvantageous manner, individual pulses are therefore not possible. This also has the disadvantage that too high a "dose" of cavitation events act on the cells and the cells are thereby destroyed.
  • the considerations that led to the creation of the invention adopted the knowledge that a hollow cylindrical device into which a multitude of in-phase sound pulses are introduced radially from the outside produces a reproducible zone of transient cavitation events in the area around the axis of rotation of the hollow cylinder.
  • the considerations that led to the development of the invention assumed that it is irrelevant for the transfer of the molecules into the target cells, how large is the total amount of the medium in which the molecules and target cells to be transferred are located, if any parts from the total amount successively reach the effective range of the acoustic pulses and the target cells contained therein are thus exposed to treatment.
  • This relative movement can now take place by moving the source relative to a fixed liquid container or by moving the liquid container relative to a fixed source.
  • the medium with the target cells and the molecules to be transferred can also move through a line system while it is under the influence of the focused acoustic pulses.
  • the parameters must be matched to one another in such a way that the target cells are exposed to at least the predetermined number of pulses as they move through the line system.
  • FIG. 1 shows a possible embodiment of the invention.
  • Acoustic pulses are generated by means of an essentially hollow cylindrical source 1, through which a tubular line 2 is guided.
  • the medium is located in this line 2.
  • it is a liquid F that has a viscosity that allows it to flow through line 2.
  • the source 1 is designed in such a way that it focuses the acoustic pulses in its interior and thus in the area in which the line 2 runs.
  • the focus here is essentially axial.
  • the area of the focus is the so-called effective area of the acoustic pulses, which is why it is called the focus area in the following.
  • the source 1 is designed to be rotationally symmetrical cylindrical.
  • the wall of line 2 is permeable to acoustic pulses at least in the area of the focus.
  • the principle of excitation for the acoustic pulses is to use a large number of piezo elements, all of which are concentrically directed in the direction of the axis of rotation and are excited simultaneously, in phase. In this way it is possible to create an elongated focus area.
  • cylindrical source instead of a plurality of piezo elements concentrically aligned with the axis of rotation by means of a plurality of piezo rings lying next to one another, the center of which lie on the axis of rotation. It is also possible to use magnetostrictive actuators to generate the acoustic pulses
  • a further embodiment of the invention provides for the source 1 to be designed as a hollow body with a semicircular cross section as a more channel-shaped body.
  • the source 1 there is a focus area that extends approximately over the length of the source.
  • the line in which the liquid F is located runs through this elongated focus area. This has the advantage that the source can be coupled to an existing line and the line does not first have to be laid through the hollow cylindrical source 1.
  • the source can advantageously be coupled to an existing line.
  • the design of a source 1 according to FIG. 2 with a semicircular cross section offers the advantage that the source can easily be attached to a line from the side without the line having to be laid through the interior of the source.
  • a coil can also be used for the excitation, which acts on a membrane located within the coil and thus generates an inwardly directed acoustic pulse.
  • Acoustic pulses are emitted by the source 1 onto the liquid F and briefly create conditions there (pressure or negative pressure with sufficient intensity, cavitation) in order to bring about a transfer of the molecules into the target cells.
  • the molecules can be transferred into the cells within wide pressure or negative pressure ranges.
  • the pressure ranges are between 10 Mpa and 150 Mpa.
  • the vacuum ranges from -5 Mpa to -50 Mpa.
  • the intensity is between 0.5 mJ / mm2 and 5.0 mJ / mm2.
  • the flow rate Vf of the liquid F can be changed by a pump, which is not shown for reasons of clarity.
  • the source 1 is excited by a unit 3 in pulse form.
  • the target cells and the molecules to be transferred are exposed to a predetermined number of acoustic pulses of a predetermined intensity.
  • the liquid F flow through the line 2 in a pulsed or clocked manner.
  • the liquid F is always conveyed in such a way that the area which was just in front of the source 1 in the line is conveyed so far that it then comes to a standstill at the end of the source 1. While the liquid area within source 1 is in the focus of the acoustic pulses, a predetermined number becomes emitted by acoustic pulses that stimulate the transfer of molecules into the target cells.
  • the acoustic pulses that act on the liquid or generally formulated on the medium can consist of ultrasound pulses or one or more successive shock waves.
  • the intensity (unit of measurement mJ / mm2) of the ultrasound impulses or shock waves also influences both the number of intracellularly transferred molecules and the damage to the cells. With increasing intensity, there is an increase in both the number of intracellularly transferred molecules and the damaged cell.
  • the container can be of any shape here, but it is crucial that the movement of the container takes place in such a way that the entire volume of the container has successively reached the effective range of the acoustic pulses.
  • All movements of the medium relative to the source of acoustic pulses can be continuous or clocked.
  • the molecules to be transferred are fed to the medium in which the target cells are located only directly before the focus area.
  • the source shown in FIG. 3 consists of an inner ring 3, which can accommodate the line (not shown here) or a sample tube in its inner free space 7.
  • Piezo elements 4 are arranged distributed around the outer circumference of the inner ring 3. For reasons of clarity, only some of these piezo elements 4 are shown in this illustration.
  • the piezo elements 4 are held on their side facing away from the inner ring 3 by a further ring 6.
  • the outer ring 6 is designed as a clamping ring. Its inside diameter can be changed using a clamping screw 2. This also changes the gap 1.
  • the piezo elements 4 can be firmly clamped between the inner ring 3 and the outer clamping ring 6. This ensures good sound transmission between the piezo elements 4 and the rings 4 and 6.
  • FIG. 4 shows a further version of the device according to the invention.
  • the piezo elements 4 are arranged around the inner ring 3 in the manner known from FIG. 3.
  • An intermediate ring 8 rests on its side facing away from the inner ring 3.
  • Piezo elements 7 are in turn arranged on this intermediate ring 8. These piezo elements 7 are followed by the clamping ring 6 already known from FIG. 3.

Landscapes

  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Sustainable Development (AREA)
  • Microbiology (AREA)
  • Biomedical Technology (AREA)
  • Cell Biology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Mechanical Engineering (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

L'invention concerne un milieu dans lequel se trouvent des cellules et des molécules à transférer dans ces cellules. Ledit milieu est remué dans un dispositif cylindrique creux, afin de produire des impulsions acoustiques induisant une cavitation, par rapport à une zone où les impulsions acoustiques interviennent de manière focalisée.
EP00990583A 1999-12-23 2000-12-23 Procede pour transferer des molecules dans des cellules Withdrawn EP1244770A2 (fr)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
DE1999162904 DE19962904A1 (de) 1999-12-23 1999-12-23 Verfahren zum Transfer von Molekülen in Zellen und Vorrichtung zur Durchführung des Verfahrens
DE19962904 1999-12-23
DE10063942 2000-12-20
DE10063942 2000-12-20
PCT/DE2000/004631 WO2001048181A2 (fr) 1999-12-23 2000-12-23 Procede pour transferer des molecules dans des cellules

Publications (1)

Publication Number Publication Date
EP1244770A2 true EP1244770A2 (fr) 2002-10-02

Family

ID=26008022

Family Applications (1)

Application Number Title Priority Date Filing Date
EP00990583A Withdrawn EP1244770A2 (fr) 1999-12-23 2000-12-23 Procede pour transferer des molecules dans des cellules

Country Status (5)

Country Link
US (1) US20030017578A1 (fr)
EP (1) EP1244770A2 (fr)
JP (1) JP2003533974A (fr)
CA (1) CA2397271A1 (fr)
WO (1) WO2001048181A2 (fr)

Families Citing this family (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE10108799A1 (de) * 2001-02-19 2002-09-05 Laser & Med Tech Gmbh Verfahren und Vorrichtung zur Ultraschallimpfung von biologischem Zellmaterial
DE10211886B4 (de) * 2002-03-18 2004-07-15 Dornier Medtech Gmbh Verfahren und Einrichtung zum Erzeugen bipolarer akustischer Impulse
DE10223196B4 (de) * 2002-05-24 2004-05-13 Dornier Medtech Systems Gmbh Verfahren und Einrichtung zum Transferieren von Molekülen in Zellen
IL149932A0 (en) * 2002-05-30 2002-11-10 Nano Size Ltd High power ultrasonic reactor and process for ultrasonic treatment of a reaction material
US7824901B2 (en) * 2003-12-01 2010-11-02 Cyto Pulse Sciences, Inc. Non-uniform electric field chamber for cell fusion
DE602005019367D1 (de) 2004-12-15 2010-04-01 Dornier Medtech Systems Gmbh Verbesserte Zelltherapie und Gewebsregeneration mittels Stosswellen bei Patienten mit kardiovaskulären and neurologischen Krankheiten
FR2957532B1 (fr) * 2010-03-19 2012-09-28 Commissariat Energie Atomique Agitateur d'un echantillon liquide
US11046596B2 (en) 2012-10-25 2021-06-29 Hydrus Technology Pty. Ltd. Electrochemical liquid treatment apparatus
US11046595B2 (en) 2014-05-23 2021-06-29 Hydrus Technology Pty. Ltd. Electrochemical treatment methods
CA2949865C (fr) 2014-05-23 2023-10-24 Hydrus Technology Pty. Ltd. Procedes de traitement electrochimique
EP3242946A4 (fr) * 2015-01-07 2018-09-26 Indee. Inc. Procédé de transfection microfluidique mécanique et hydrodynamique et appareil correspondant
KR102232757B1 (ko) * 2018-11-22 2021-03-26 (주)엑솔런스바이오테크놀로지 체외충격파를 이용한 표적물질 전달 장치

Family Cites Families (11)

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Publication number Priority date Publication date Assignee Title
US2578505A (en) * 1948-03-02 1951-12-11 Sperry Prod Inc Supersonic agitation
US3406302A (en) * 1966-03-15 1968-10-15 Westinghouse Electric Corp Cylindrical magnetostrictive electromechanical transducer
DE2346649A1 (de) * 1973-09-17 1975-03-27 Ngk Spark Plug Co Ultraschallgeber
US4369100A (en) * 1977-09-27 1983-01-18 Sawyer Harold T Method for enhancing chemical reactions
DD284131A7 (de) * 1987-07-10 1990-11-07 �����@�������`����k�� Verfahren und vorrichtung zur herstellung bioaktiver suspensionen
GB8721015D0 (en) * 1987-09-07 1987-10-14 Amersham Int Plc Modifying living cells
DE3803275A1 (de) * 1988-02-04 1989-08-17 Dornier Medizintechnik Piezoelektrische stosswellenquelle
US5395592A (en) * 1993-10-04 1995-03-07 Bolleman; Brent Acoustic liquid processing device
DE19820466C2 (de) * 1998-05-07 2002-06-13 Fraunhofer Ges Forschung Vorrichtung und Verfahren zur gezielten Beaufschlagung einer biologischen Probe mit Schallwellen
CA2238951A1 (fr) * 1998-05-26 1999-11-26 Les Technologies Sonomax Inc. Reacteur a cavitation acoustique pour le traitement des materiaux
DE19834612A1 (de) * 1998-07-31 2000-02-24 Dornier Medtech Holding Int Gmbh Verfahren zum intrazellulären Transfer von Oligonukleotiden und Vorrichtung zur Durchführung desselben

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO0148181A2 *

Also Published As

Publication number Publication date
WO2001048181A3 (fr) 2002-04-18
WO2001048181A2 (fr) 2001-07-05
JP2003533974A (ja) 2003-11-18
CA2397271A1 (fr) 2001-07-05
US20030017578A1 (en) 2003-01-23

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