WO2022109113A1 - Methods and systems for mechanoporation-based payload delivery into biological cells - Google Patents

Methods and systems for mechanoporation-based payload delivery into biological cells Download PDF

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Publication number
WO2022109113A1
WO2022109113A1 PCT/US2021/059856 US2021059856W WO2022109113A1 WO 2022109113 A1 WO2022109113 A1 WO 2022109113A1 US 2021059856 W US2021059856 W US 2021059856W WO 2022109113 A1 WO2022109113 A1 WO 2022109113A1
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WO
WIPO (PCT)
Prior art keywords
cell
processing
processing apparatus
cells
inlet
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.)
Ceased
Application number
PCT/US2021/059856
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English (en)
French (fr)
Inventor
Sewoon HAN
Ian SICHER
Alexander Alexeev
Ockchul KIM
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.)
CellFE Inc
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CellFE Inc
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 to IL302976A priority Critical patent/IL302976A/en
Priority to JP2023529914A priority patent/JP7642813B2/ja
Priority to EP21895569.8A priority patent/EP4232554A1/en
Priority to KR1020237020303A priority patent/KR102819986B1/ko
Priority to AU2021383756A priority patent/AU2021383756A1/en
Priority to CN202180090664.XA priority patent/CN117015596B/zh
Application filed by CellFE Inc filed Critical CellFE Inc
Priority to CA3201944A priority patent/CA3201944A1/en
Publication of WO2022109113A1 publication Critical patent/WO2022109113A1/en
Priority to US18/162,372 priority patent/US11788050B2/en
Anticipated expiration legal-status Critical
Priority to US18/456,340 priority patent/US12359158B2/en
Ceased legal-status Critical Current

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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
    • 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
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/04—Filters; Permeable or porous membranes or plates, e.g. dialysis
    • 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
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/0062—General methods for three-dimensional culture
    • 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
    • C12N2527/00—Culture process characterised by the use of mechanical forces, e.g. strain, vibration

Definitions

  • FIG. 4B is a schematic exploded view of the cell processing apparatus in FIG. 4A.
  • Microfluidic techniques provide new opportunities for processing and manipulation of biological cells, such as the delivery of payload into cells for gene engineering and other applications.
  • a “microfluidic” technique is defined as a process of passing fluid through a channel, which has the smallest dimension of less than 1 millimeter.
  • an apparatus may include one or more constrictions forming a gap that is less than 1 millimeter.
  • microfluidic techniques are the small size of flow channels, resulting in flow conditions represented by low Reynolds numbers (e.g., less than 1). As such, fluid viscosity is a dominant factor of these flow conditions. Furthermore, the overall throughput of a single channel is limited. For example, a volumetric flow rate of a single channel having a width of 1 millimeter can be less than 1 ml/min. This flow rate corresponds to the processing speed of about 10 7 cells per hour. At the same time, industrial applications require much larger processing speeds, such as 10 9 or 10 10 cells per hour, which has significantly limited the adoption of microfluidic techniques in the past.
  • FIG. 1A and FIG. IB are schematic representations of cell processing apparatus 100 (in an assembled form and exploded view), in accordance with some examples.
  • Cell processing apparatus 100 comprises processing assembly 110, inlet component 102, and stopper 104.
  • Processing assembly 110 is disposed between inlet component 102 and stopper 104. More specifically, each inlet component 102 and stopper 104 is sealed against processing assembly 110.
  • processing assembly 110, inlet component 102, and stopper 104 define and enclose cavity 107 of cell processing apparatus 100. Cavity 107 can be also referred to as an inlet opening.
  • cell processing apparatus 100 also comprises distribution component 106, positioned inside cavity 107.
  • ridges 140 are clustered closer to the channel inlet, e.g., to allow longer time for cells to remain in the fluid with higher speed after interactions with ridges 140.
  • the channel inlet and/or the channel outlet are located at the front and back edges of processing components 119.
  • the channel inlet and/or outlet can be arranged through channel wall 132 of processing component 119, e.g., to reduce the amount of dust entering each channel 130 due to the fabrication process.

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  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Chemical & Material Sciences (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Sustainable Development (AREA)
  • Cell Biology (AREA)
  • Mechanical Engineering (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Molecular Biology (AREA)
  • Plant Pathology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
PCT/US2021/059856 2020-11-18 2021-11-18 Methods and systems for mechanoporation-based payload delivery into biological cells Ceased WO2022109113A1 (en)

Priority Applications (9)

Application Number Priority Date Filing Date Title
JP2023529914A JP7642813B2 (ja) 2020-11-18 2021-11-18 生体細胞へのメカノポレーションベースのペイロード送達のための方法及びシステム
EP21895569.8A EP4232554A1 (en) 2020-11-18 2021-11-18 Methods and systems for mechanoporation-based payload delivery into biological cells
KR1020237020303A KR102819986B1 (ko) 2020-11-18 2021-11-18 생물학적 세포로 기계적 천공 기반 페이로드 전달을 위한 방법 및 시스템
AU2021383756A AU2021383756A1 (en) 2020-11-18 2021-11-18 Methods and systems for mechanoporation-based payload delivery into biological cells
CN202180090664.XA CN117015596B (zh) 2020-11-18 2021-11-18 机械穿孔类有效负载递送至生物细胞的方法和系统
IL302976A IL302976A (en) 2020-11-18 2021-11-18 Mechanoporation-based methods and systems for high-throughput charge transfer into biological cells
CA3201944A CA3201944A1 (en) 2020-11-18 2021-11-18 Methods and systems for mechanoporation-based payload delivery into biological cells
US18/162,372 US11788050B2 (en) 2020-11-18 2023-01-31 Methods and systems for mechanoporation-based high-throughput payload delivery into biological cells
US18/456,340 US12359158B2 (en) 2020-11-18 2023-08-25 Methods and systems for mechanoporation-based high-throughput payload delivery into biological cells

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US202063115507P 2020-11-18 2020-11-18
US63/115,507 2020-11-18

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US18/162,372 Continuation US11788050B2 (en) 2020-11-18 2023-01-31 Methods and systems for mechanoporation-based high-throughput payload delivery into biological cells

Publications (1)

Publication Number Publication Date
WO2022109113A1 true WO2022109113A1 (en) 2022-05-27

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PCT/US2021/059856 Ceased WO2022109113A1 (en) 2020-11-18 2021-11-18 Methods and systems for mechanoporation-based payload delivery into biological cells

Country Status (9)

Country Link
US (2) US11788050B2 (cg-RX-API-DMAC7.html)
EP (1) EP4232554A1 (cg-RX-API-DMAC7.html)
JP (1) JP7642813B2 (cg-RX-API-DMAC7.html)
KR (1) KR102819986B1 (cg-RX-API-DMAC7.html)
CN (1) CN117015596B (cg-RX-API-DMAC7.html)
AU (1) AU2021383756A1 (cg-RX-API-DMAC7.html)
CA (1) CA3201944A1 (cg-RX-API-DMAC7.html)
IL (1) IL302976A (cg-RX-API-DMAC7.html)
WO (1) WO2022109113A1 (cg-RX-API-DMAC7.html)

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CN115254214A (zh) * 2022-06-29 2022-11-01 中国科学院精密测量科学与技术创新研究院 一种微流控通道、微流控芯片和生化分子递送方法
US11788050B2 (en) 2020-11-18 2023-10-17 Cellfe, Inc. Methods and systems for mechanoporation-based high-throughput payload delivery into biological cells
US12227729B2 (en) 2020-12-24 2025-02-18 Cellfe, Inc. Methods and systems for high-throughput cell processing
WO2025194040A1 (en) * 2024-03-15 2025-09-18 Cellfe, Inc. High-throughput mechanoporation for scalable and cost-effective cell therapy manufacturing

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Publication number Priority date Publication date Assignee Title
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WO2025194040A1 (en) * 2024-03-15 2025-09-18 Cellfe, Inc. High-throughput mechanoporation for scalable and cost-effective cell therapy manufacturing

Also Published As

Publication number Publication date
US20230416781A1 (en) 2023-12-28
IL302976A (en) 2023-07-01
US20230174918A1 (en) 2023-06-08
JP2023550090A (ja) 2023-11-30
US12359158B2 (en) 2025-07-15
EP4232554A1 (en) 2023-08-30
JP7642813B2 (ja) 2025-03-10
KR102819986B1 (ko) 2025-06-12
KR20230110308A (ko) 2023-07-21
US11788050B2 (en) 2023-10-17
AU2021383756A1 (en) 2023-06-22
CA3201944A1 (en) 2022-05-27
CN117015596B (zh) 2024-02-09
CN117015596A (zh) 2023-11-07

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