WO2004044127A2 - Populations de cellules souches - Google Patents

Populations de cellules souches Download PDF

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Publication number
WO2004044127A2
WO2004044127A2 PCT/US2003/033368 US0333368W WO2004044127A2 WO 2004044127 A2 WO2004044127 A2 WO 2004044127A2 US 0333368 W US0333368 W US 0333368W WO 2004044127 A2 WO2004044127 A2 WO 2004044127A2
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WO
WIPO (PCT)
Prior art keywords
cells
population
tgf
positive
beta
Prior art date
Application number
PCT/US2003/033368
Other languages
English (en)
Other versions
WO2004044127A3 (fr
Inventor
Morey Kraus
Original Assignee
Viacell, 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
Application filed by Viacell, Inc. filed Critical Viacell, Inc.
Priority to AU2003284314A priority Critical patent/AU2003284314A1/en
Publication of WO2004044127A2 publication Critical patent/WO2004044127A2/fr
Publication of WO2004044127A3 publication Critical patent/WO2004044127A3/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
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0634—Cells from the blood or the immune system
    • C12N5/0647—Haematopoietic stem cells; Uncommitted or multipotent progenitors

Definitions

  • This invention relates to stem cell populations and methods of obtaining such populations.
  • TGF-beta plays a role in controlling cell cycles in many human cell types.
  • TGF-beta has diverse functionality, which includes both proliferative and differentiative aspects.
  • TGF-beta functions differently in different cell types. In some cases, it plays a role in stimulating cells to grow. In other cases, it plays an opposite role, maintaining the cells in a state of quiescence (a non-cycling state).
  • the dominant role of TGF-beta in the most primitive of hematopoietic stem cells is to maintain quiescence. Early hematopoietic stem cells bear TGF-beta receptors.
  • TGF-beta production methods of the invention do not interfere, interact, or otherwise disturb TGF-beta receptors on the surface of these target cells.
  • Enrichment is enhanced by lineage depletion (using positive or negative selection to remove Lin+ cells, using, e.g., an antibody) as is described in Kraus U.S. Patent No. 5,925,567, the disclosure of which is hereby incorporated herein by reference.
  • methods of the invention yield enriched stem cell containing populations, which can be expanded, and can be of value to patients in need of cellular therapy, e.g., cancer therapy, immunotherapy, and gene therapy.
  • the invention features a population of human cells containing at least 1% TGF-beta positive/Lineage depleted cells.
  • the TGF-beta positive/Lineage depleted cells can be derived from human cord blood, bone marrow or other sources. At least 5%, preferably at least 50%, and more preferably at least 90% of the cells in the population are TGF-beta positive/Lineage depleted cells.
  • the cell population may be further stem cell enriched by selecting cells that are also CD45+/CD34+/CD38-.
  • Lineage positive cells can be removed from the starting population of relatively undifferentiated cells by negative selection.
  • Target cells can be expanded following selection, e.g., by the methods described in U.S. Patent No. 5,925,567.
  • TGF- ⁇ positive/Lineage depleted cells can again be removed from the target cell population to further purify the target cell population.
  • the removing step can be performed using flow cytometry, e.g., using fluorescent activated cell sorting.
  • the method includes (a) selecting TGF-beta positive cells by positive selection; and (b) depleting TGF-beta positive cells of Lineage minus cells by negative selection.
  • steps (a) and (b) are performed sequentially.
  • the invention further features methods of treating a patient in need of cellular therapy by administering to the patient an aliquot of one of the cell populations described above.
  • Fig. 1 is a dot plot showing the population profile of a Human Cord Blood Lineage Negative (CD2/3/4/8/14/16/19/24/56/66b/GlyA) Cell Population that includes a subpopulation (0.16%) which is co-positive for 1D11.
  • Fig. 2 is a dot plot showing the population of Fig. 1, back-gated into a forward/side scatter plot, indicating that 67% of the Lin-/1D11+ cells fall within the primitive leucocyte gate.
  • Figs. 3 and 3 A are dot plots showing the presence of mouse Lin- /Rholo/Holo/c-Kit+Sca+ hematopoietic stem cells (Fig. 3 A) in lineage-depleted bone marrow cell preparations (Fig. 3).
  • Isolation and enrichment can be performed using a variety of immuno- selection strategies, including panning, magnetic particles, magnetic beads, chromatographic-like techniques, cell sorting, and high-speed cell sorting.
  • suitable strategies include flow cytometry and fluorescent activated cell sorting.
  • This population can be expanded, e.g., using the target cell expansion methods described in U.S. Patent No. 5,925,567, the disclosure of which is incorporated herein by reference.
  • HCB obtained from full term infants was red cell depleted according to standard procedures and cryopreserved.
  • the processed cord was later thawed and lineage depleted by immunomagnetic means using a cocktail of lineage antibodies that included CD2/CD3/CD14/CD16/CD19/CD24/CD56/CD66b/GlyA (StemCell Technologies).
  • a sample of the purified fraction was incubated with 80 ug/ml of anti-TGF-beta antibody (IDl 1) that had been conjugated with ALEXA-488 (Molecular Probes) fluorochrome.
  • IDl 1 anti-TGF-beta antibody
  • ALEXA-488 Molecular Probes fluorochrome
  • the purified anti-TGF-beta positive/LIN- populations of the invention are separated from lineage depleted HCB using a high speed cell sorter (e.g., Cytomation). These cells can be functionally characterized in CFC, HPP and NOD/scid engraftrnent assays.
  • the method represents a 100 to 1,000 fold enrichment in the target population.
  • Other embodiments are within the scope of the following claims.

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  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biomedical Technology (AREA)
  • Genetics & Genomics (AREA)
  • Zoology (AREA)
  • Hematology (AREA)
  • Biotechnology (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical & Material Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Microbiology (AREA)
  • Immunology (AREA)
  • Developmental Biology & Embryology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Cell Biology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

La présente invention concerne des populations de cellules humaines comprenant au moins 1 % de cellules affaiblie TGF-bêta positives/de lignée. Cette invention concerne aussi des techniques permettant d'obtenir ces populations.
PCT/US2003/033368 2002-11-08 2003-10-20 Populations de cellules souches WO2004044127A2 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2003284314A AU2003284314A1 (en) 2002-11-08 2003-10-20 Stem cell populations

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US42483502P 2002-11-08 2002-11-08
US60/424,835 2002-11-08

Publications (2)

Publication Number Publication Date
WO2004044127A2 true WO2004044127A2 (fr) 2004-05-27
WO2004044127A3 WO2004044127A3 (fr) 2004-07-08

Family

ID=32312880

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2003/033368 WO2004044127A2 (fr) 2002-11-08 2003-10-20 Populations de cellules souches

Country Status (2)

Country Link
AU (1) AU2003284314A1 (fr)
WO (1) WO2004044127A2 (fr)

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11608486B2 (en) 2015-07-02 2023-03-21 Terumo Bct, Inc. Cell growth with mechanical stimuli
US11613727B2 (en) 2010-10-08 2023-03-28 Terumo Bct, Inc. Configurable methods and systems of growing and harvesting cells in a hollow fiber bioreactor system
US11624046B2 (en) 2017-03-31 2023-04-11 Terumo Bct, Inc. Cell expansion
US11629332B2 (en) 2017-03-31 2023-04-18 Terumo Bct, Inc. Cell expansion
US11634677B2 (en) 2016-06-07 2023-04-25 Terumo Bct, Inc. Coating a bioreactor in a cell expansion system
US11667876B2 (en) 2013-11-16 2023-06-06 Terumo Bct, Inc. Expanding cells in a bioreactor
US11667881B2 (en) 2014-09-26 2023-06-06 Terumo Bct, Inc. Scheduled feed
US11685883B2 (en) 2016-06-07 2023-06-27 Terumo Bct, Inc. Methods and systems for coating a cell growth surface
US11795432B2 (en) 2014-03-25 2023-10-24 Terumo Bct, Inc. Passive replacement of media
US11965175B2 (en) 2016-05-25 2024-04-23 Terumo Bct, Inc. Cell expansion

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5925567A (en) * 1995-05-19 1999-07-20 T. Breeders, Inc. Selective expansion of target cell populations

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5925567A (en) * 1995-05-19 1999-07-20 T. Breeders, Inc. Selective expansion of target cell populations

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
BATARD, ET AL: 'TGF-BETA 1 MAINTAINS HEMATOPOIETIC IMMATURITY BY A REVERSIBLE NEGATIVE CONTROL OF CELL CYCLE AND INDUCES CD34 ANTIGEN UP-MODULATION' JOURNAL OF CELL SCIENCE vol. 113, 2000, pages 383 - 390, XP000939356 *
VITOLO D, ET AL: 'RAPID IL-2-INDUCED ADHERENCE OF HUMAN NATURAL KILLER CELLS' THE JOURNAL OF IMMUNOLOGY vol. 151, no. 4, August 1993, pages 1926 - 1937, XP002977043 *

Cited By (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11613727B2 (en) 2010-10-08 2023-03-28 Terumo Bct, Inc. Configurable methods and systems of growing and harvesting cells in a hollow fiber bioreactor system
US11773363B2 (en) 2010-10-08 2023-10-03 Terumo Bct, Inc. Configurable methods and systems of growing and harvesting cells in a hollow fiber bioreactor system
US11746319B2 (en) 2010-10-08 2023-09-05 Terumo Bct, Inc. Customizable methods and systems of growing and harvesting cells in a hollow fiber bioreactor system
US11708554B2 (en) 2013-11-16 2023-07-25 Terumo Bct, Inc. Expanding cells in a bioreactor
US11667876B2 (en) 2013-11-16 2023-06-06 Terumo Bct, Inc. Expanding cells in a bioreactor
US11795432B2 (en) 2014-03-25 2023-10-24 Terumo Bct, Inc. Passive replacement of media
US11667881B2 (en) 2014-09-26 2023-06-06 Terumo Bct, Inc. Scheduled feed
US11608486B2 (en) 2015-07-02 2023-03-21 Terumo Bct, Inc. Cell growth with mechanical stimuli
US11965175B2 (en) 2016-05-25 2024-04-23 Terumo Bct, Inc. Cell expansion
US11685883B2 (en) 2016-06-07 2023-06-27 Terumo Bct, Inc. Methods and systems for coating a cell growth surface
US11634677B2 (en) 2016-06-07 2023-04-25 Terumo Bct, Inc. Coating a bioreactor in a cell expansion system
US11999929B2 (en) 2016-06-07 2024-06-04 Terumo Bct, Inc. Methods and systems for coating a cell growth surface
US11702634B2 (en) 2017-03-31 2023-07-18 Terumo Bct, Inc. Expanding cells in a bioreactor
US11629332B2 (en) 2017-03-31 2023-04-18 Terumo Bct, Inc. Cell expansion
US11624046B2 (en) 2017-03-31 2023-04-11 Terumo Bct, Inc. Cell expansion

Also Published As

Publication number Publication date
WO2004044127A3 (fr) 2004-07-08
AU2003284314A1 (en) 2004-06-03
AU2003284314A8 (en) 2004-06-03

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