WO1988004322A1 - Injection moleculaire cellulaire a gravite artificielle - Google Patents

Injection moleculaire cellulaire a gravite artificielle Download PDF

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Publication number
WO1988004322A1
WO1988004322A1 PCT/US1987/003212 US8703212W WO8804322A1 WO 1988004322 A1 WO1988004322 A1 WO 1988004322A1 US 8703212 W US8703212 W US 8703212W WO 8804322 A1 WO8804322 A1 WO 8804322A1
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WO
WIPO (PCT)
Prior art keywords
cells
rotor
membrane
artificial gravity
centrifuge
Prior art date
Application number
PCT/US1987/003212
Other languages
English (en)
Inventor
Jean P. Biscar
Original Assignee
Biscar Jean P
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
Application filed by Biscar Jean P filed Critical Biscar Jean P
Publication of WO1988004322A1 publication Critical patent/WO1988004322A1/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
    • 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
    • C12M35/02—Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N13/00—Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves

Definitions

  • the extraction phase is completed, and the injection phase initiated, by suddenly, rapidly and continuously decreasing the angular velocity of the centrifuge rotor, and, hence, the artificial gravity field within the centrifuge chamber.
  • the resulting pressure differential across the cellular membranes induced thereby continuously forces the poration medium in proximity to the cells, now impregnate with molecules, through the pores in the cellular membranes.
  • the inflow of impregnated medium also maintains membrane porosity by preventing the membrane periphery of the pores from closing/healing, thereby allowing further inflow of impregnated medium into the cells.
  • the rate of inflow of impregnated medium, and, hence, the rate of molecular injection is controlled by adjustment of the angular deceleration of the centrifuge rotor.
  • FIG. 1 is a perspective view of a centrifuge constructed in accordance with the present invention.
  • a pair of diametrically opposed apertures 76 is formed in the peripheral edge of the disk 38 extending radially inward in the plane thereof with sufficient depth so as to intersect the central aperture 40 formed therein and penetrate through the outer cylinder 46 of the hub sub-assembly 34.
  • An electrically conductive rod 78 is internally disposed therein, having its ends secured to the peripheral wall 36 and the inner cylinder 42, respectively, upon assembly.
  • FIG. 4 additionally shows a plurality of contiguous alternating reflective and nonreflective surface markings 80 applied to the underside 54 of the rotor assembly 32.
  • the markings 80 are used in conjunction with the angular velocity indicator 20 through use of a digital optical speed sensor (not shown) .

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  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • General Health & Medical Sciences (AREA)
  • Cell Biology (AREA)
  • Sustainable Development (AREA)
  • Physics & Mathematics (AREA)
  • Electromagnetism (AREA)
  • Mechanical Engineering (AREA)
  • Biophysics (AREA)
  • Molecular Biology (AREA)
  • Plant Pathology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

Procédé et appareil d'introduction de molécules et de particules à l'intérieur de cellules vivantes. L'invention utilise la gravité artificielle créée par un rotor tournant. Cette gravité artificielle produit plusieurs effets: elle produit une seule couche de cellules contre la surface plate externe; elle comprime les cellules contre la surface plate, en mettant l'intérieur des cellules sous pression; puis une impulsion électrique forme une ouverture au sommet des membranes des cellules. L'accélération du rotor permet l'extraction contrôlée du cytoplasme de l'intérieur des cellules; dès qu'une quantité suffisante de cytoplasme a été extraite, le trou dans la membrane est maintenu ouvert, le rotor est ralenti, et la réduction de la gravité artificielle s'exerçant sur les cellules leur permet de reprendre leur forme originale. Pendant ce processus, les cellules aspirent, au travers du trou ouvert ménagé dans la membrane, le liquide environnant ainsi que les molécules à injecter dans les cellules. L'appareil comprend une centrifugeuse et son rotor, une unité à chambre (100) pour les cellules, cette unité étant montée sur le rotor. L'unité à chambre pour cellules comprend une chambre à cellules (140), des électrodes externe et interne (104, 124), une ouverture (130) et des passages reliant l'ouverture (130) à la chambre à cellules (140) ainsi qu'un mécanisme de freinage pour le rotor de la centrifugeuse.
PCT/US1987/003212 1986-12-07 1987-12-04 Injection moleculaire cellulaire a gravite artificielle WO1988004322A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US93860786A 1986-12-07 1986-12-07
US938,607 1986-12-07

Publications (1)

Publication Number Publication Date
WO1988004322A1 true WO1988004322A1 (fr) 1988-06-16

Family

ID=25471664

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1987/003212 WO1988004322A1 (fr) 1986-12-07 1987-12-04 Injection moleculaire cellulaire a gravite artificielle

Country Status (1)

Country Link
WO (1) WO1988004322A1 (fr)

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5612207A (en) * 1993-03-23 1997-03-18 Cbr Laboratories, Inc. Method and apparatus for encapsulation of biologically-active substances in cells
US5720921A (en) * 1995-03-10 1998-02-24 Entremed, Inc. Flow electroporation chamber and method
US6004663A (en) * 1995-03-29 1999-12-21 E. I. Du Pont De Nemours And Company Solid surface material with foam backing
US6074605A (en) * 1995-03-10 2000-06-13 Entremed, Inc. Flow electroporation chamber and method
US6582636B1 (en) 1995-03-29 2003-06-24 E. I. Du Pont De Nemours And Company Two-step process for producing solid surface material with foam backing
US6773669B1 (en) 1995-03-10 2004-08-10 Maxcyte, Inc. Flow electroporation chamber and method
US7029916B2 (en) 2001-02-21 2006-04-18 Maxcyte, Inc. Apparatus and method for flow electroporation of biological samples
US7141425B2 (en) 2001-08-22 2006-11-28 Maxcyte, Inc. Apparatus and method for electroporation of biological samples
US7771984B2 (en) 2004-05-12 2010-08-10 Maxcyte, Inc. Methods and devices related to a regulated flow electroporation chamber

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4456581A (en) * 1980-11-25 1984-06-26 Boehringer Mannheim Gmbh Centrifugal analyzer rotor unit and insert elements
US4695547A (en) * 1986-04-02 1987-09-22 Jeffrey L. Hilliard Probe for electrofusion, electroporation, or like procedure

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4456581A (en) * 1980-11-25 1984-06-26 Boehringer Mannheim Gmbh Centrifugal analyzer rotor unit and insert elements
US4695547A (en) * 1986-04-02 1987-09-22 Jeffrey L. Hilliard Probe for electrofusion, electroporation, or like procedure

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5612207A (en) * 1993-03-23 1997-03-18 Cbr Laboratories, Inc. Method and apparatus for encapsulation of biologically-active substances in cells
US5720921A (en) * 1995-03-10 1998-02-24 Entremed, Inc. Flow electroporation chamber and method
US6074605A (en) * 1995-03-10 2000-06-13 Entremed, Inc. Flow electroporation chamber and method
US6773669B1 (en) 1995-03-10 2004-08-10 Maxcyte, Inc. Flow electroporation chamber and method
US6004663A (en) * 1995-03-29 1999-12-21 E. I. Du Pont De Nemours And Company Solid surface material with foam backing
US6582636B1 (en) 1995-03-29 2003-06-24 E. I. Du Pont De Nemours And Company Two-step process for producing solid surface material with foam backing
US7029916B2 (en) 2001-02-21 2006-04-18 Maxcyte, Inc. Apparatus and method for flow electroporation of biological samples
US7141425B2 (en) 2001-08-22 2006-11-28 Maxcyte, Inc. Apparatus and method for electroporation of biological samples
US7186559B2 (en) 2001-08-22 2007-03-06 Maxcyte, Inc. Apparatus and method for electroporation of biological samples
US7771984B2 (en) 2004-05-12 2010-08-10 Maxcyte, Inc. Methods and devices related to a regulated flow electroporation chamber
US9546350B2 (en) 2004-05-12 2017-01-17 Maxcyte, Inc. Methods and devices related to a regulated flow electroporation chamber

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