EP1056835A1 - Derivation de cellules et de tissus a partir du stade cellulaire pre-souche pour les therapies de transplantation - Google Patents

Derivation de cellules et de tissus a partir du stade cellulaire pre-souche pour les therapies de transplantation

Info

Publication number
EP1056835A1
EP1056835A1 EP99909633A EP99909633A EP1056835A1 EP 1056835 A1 EP1056835 A1 EP 1056835A1 EP 99909633 A EP99909633 A EP 99909633A EP 99909633 A EP99909633 A EP 99909633A EP 1056835 A1 EP1056835 A1 EP 1056835A1
Authority
EP
European Patent Office
Prior art keywords
cells
stem cells
embryonic stem
line
embryonic
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Withdrawn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
EP99909633A
Other languages
German (de)
English (en)
Other versions
EP1056835A4 (fr
Inventor
Gary D. Hodgen
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.)
Medical College of Hampton Roads
Original Assignee
Medical College of Hampton Roads
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 Medical College of Hampton Roads filed Critical Medical College of Hampton Roads
Publication of EP1056835A1 publication Critical patent/EP1056835A1/fr
Publication of EP1056835A4 publication Critical patent/EP1056835A4/fr
Withdrawn legal-status Critical Current

Links

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
    • C12N5/0603—Embryonic cells ; Embryoid bodies
    • C12N5/0606—Pluripotent embryonic cells, e.g. embryonic stem cells [ES]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention relates to the derivation of cells and tissues from embryonic pre-stem cells for transplantation therapies .
  • This invention relates to the use of dispersed morula cells in preference to inner cell mass (ICM) from blastocysts.
  • the morula stage is the last pre-embryonic
  • pre-stem cells embryonic stem cells
  • the ICM from the blastocyst is already differentiated from trophoblastic cells, which are by then destined to become part of the placenta.
  • This invention also relates to the use of chimeric introductions into pre-stem cell cultures and stem cell propagations in culture. That is, "teacher-cells” or spent media from them, that derived from other sources (e.g. adults, cord blood, fetal tissues, etc.) will "teach" undifferentiated pre-stem cells how to convert to our sought-after therapeutic cell population both more rapidly and more preferentially.
  • This invention also relates to the identification and use of certain early markers of stem cell
  • the present invention provides for a method of isolating and propagating a line of embryonic stem cells that originates from either morulae (pre-stem) or blastocyst (ICM stem cells) . Therefore, Morula stage, undifferentiated pre-stem cells will be used as progenitors of stem cell populations. The propagated line of embryonic stem cells are then used for the purpose of transplanting cells, tissues or organs.
  • the propagation of stem cells can be initiated by formation of chimeric inner cell mass cells.
  • Chimeric ICMs will be developed from blastocysts. From such ICMs, superior stem cell cultures are derived.
  • the formation of chimeric inner cell mass cells comprises nuclear transplantation, mitochondrial substitution, or cytoplasmic depletion.
  • At least one regulatory factor is used to propagate the line of embryonic stem cells. More preferably, the regulatory factor is derived from "Teacher cells” or “Teacher cells'” spent culture medium. "Teacher cells” will be introduced into less differentiated pre-stem or early stage stem cells to accelerate propagation of target stem cells. Alternatively, spent media from
  • teacher cells can be used to accelerate the propagation of the target stem cells.
  • the embryonic stem cells are cultured in a medium in the presence of at least one agent or cytokine in order to differentiate into target specific cells or tissues.
  • the agent or cytokine is selected from the group consisting of IL-1, TNF- ⁇ , IL-6, PTH, PDGF, PGE ⁇ cAMP, estrogens, anti- estrogens, progestins, anti-progestins, cortisol, GH, androgens, I 3 /T 3 , VGEF and cyclosporin.
  • the concentration of the agent or cytokine in culture medium is from about 1.0 pg/ml to about 10.0 ng/ml.
  • the target specific cells are selected from the group consisting of nerve cells, bone cells, immune cells, and pancreatic beta cells.
  • Some such markers are, for example: Fe++ sequestration, Hg accumulation, myeloid fibers, nerve growth factor, apoptotic factors, insulin synthesis, dopamine loading, hemoglobin loading, etc. Additionally, other early markers of embryonic stem cells can be identified.
  • embryonic stem (ES) cells are derived from either morula or blastocyst stage embryos by placing cells on fibroblast feeder layers. The colonies are evaluated for differentiation state using accepted markers. Further - 4 -
  • Clonal properties of the propagated stem cells can be achieved by adding apoptotic factors, cytokines or other agents to the culture medium to eliminate contaminating members of the stem cells that did not properly differentiate.
  • the cytokines or agents are selected from the group consisting of IL-1, TNF- , I -6, PTH, PDGF, PGE 2 , cAMP, estrogens, anti-estrogens, progestins, anti-progestins, cortisol, GH, androgens, I 3 /T 3 , VGEF and cyclosporin.
  • the propagation of the line of embryonic stem cells is done in vivo by transplanting "Teacher cells" into an area sufficiently close to the embryonic stem cells to allow for at least one regulatory factor made by the teacher cells to contact the embryonic cells.
  • the presence or absence of different concentrations of calcium can be used to regulate the propagation of the line of embryonic stem cells.
  • the propagated line of embryonic stem cells is grown in a three dimensional manner before being used for transplantation.
  • This invention will allow for the efficient, safe and commercially viable derivation of cells and tissues - 5 -
  • embryonic pre-stem cells for transplantation therapies. Specifically, growing-out of human blastocysts at a rate greater than 50% from the 2-cell stage of the pre-embryo should be achieved. Also, efficient harvesting of either morula stage pre-stem cells and/or stem cells isolated from the inner cell mass of blastocysts should be achieved. These embryonic pre-stem and stem cell populations should preferably remain viable in culture for more than one week.
  • Pluripotent stem cells will be isolated and directed to differentiate into hemopoietic destinies. Therefore, tissues derived from the blood cell group or beta cells of the immune response system will be replaced in deficient patients suffering from conditions such as HIV infection, post-chemotherapy, or irradiation depletion. Culture condition in vitro will direct the rate and degree of differentiation manifested by these pluripotent stem cells, such as the presence of "teacher cells” or certain additives to the media, e.g. cytokines.
  • stem cells will be modified by formation of chimeric cell lines that incorporate "hybrid" metabolic functions that when transplanted will provide the transplant recipient with long-term relief from organ/tissue deficiencies. For instance, the production of dopamine in situ can modify neurological treatments for patients manifesting muscular rigidity and loss of motor control in disease states such as Parkinson's disease. Unlike pharmaceutical therapeutics which are partially effective temporarily, transplantation of chimeric stem cells that regulate the production dopamine and serotonergic factors will offer these patients superior outcomes.

Landscapes

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

Abstract

L'invention concerne un procédé qui permet d'isoler et de propager une lignée de cellules souches embryonnaires à partir du stade morula (pré-souche) ou blastocyste (souche à masse cellulaire interne (ICM)), pour les besoins des transplantations de cellules, de tissus ou d'organes.
EP99909633A 1998-02-27 1999-02-26 Derivation de cellules et de tissus a partir du stade cellulaire pre-souche pour les therapies de transplantation Withdrawn EP1056835A4 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US7627398P 1998-02-27 1998-02-27
US76273P 1998-02-27
PCT/US1999/004188 WO1999043785A1 (fr) 1998-02-27 1999-02-26 Derivation de cellules et de tissus a partir du stade cellulaire pre-souche pour les therapies de transplantation

Publications (2)

Publication Number Publication Date
EP1056835A1 true EP1056835A1 (fr) 2000-12-06
EP1056835A4 EP1056835A4 (fr) 2003-01-15

Family

ID=22130951

Family Applications (1)

Application Number Title Priority Date Filing Date
EP99909633A Withdrawn EP1056835A4 (fr) 1998-02-27 1999-02-26 Derivation de cellules et de tissus a partir du stade cellulaire pre-souche pour les therapies de transplantation

Country Status (5)

Country Link
EP (1) EP1056835A4 (fr)
JP (1) JP2002504362A (fr)
AU (1) AU757036B2 (fr)
CA (1) CA2320423A1 (fr)
WO (1) WO1999043785A1 (fr)

Families Citing this family (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20030113303A1 (en) * 1998-02-05 2003-06-19 Yitzhack Schwartz Homing of embryonic stem cells to a target zone in tissue using active therapeutics or substances
US7410798B2 (en) 2001-01-10 2008-08-12 Geron Corporation Culture system for rapid expansion of human embryonic stem cells
US6667176B1 (en) 2000-01-11 2003-12-23 Geron Corporation cDNA libraries reflecting gene expression during growth and differentiation of human pluripotent stem cells
US20050042749A1 (en) 2001-05-16 2005-02-24 Carpenter Melissa K. Dopaminergic neurons and proliferation-competent precursor cells for treating Parkinson's disease
US7455983B2 (en) 2000-01-11 2008-11-25 Geron Corporation Medium for growing human embryonic stem cells
WO2001088104A2 (fr) 2000-05-17 2001-11-22 Geron Corporation Populations de progeniteurs neuronaux
US7250294B2 (en) 2000-05-17 2007-07-31 Geron Corporation Screening small molecule drugs using neural cells differentiated from human embryonic stem cells
AU2002246132A1 (en) * 2002-03-13 2003-09-22 Fundacion Ivi Para El Estudio De La Reproduccion Humana (Fivier) Method of producing cell lines
CN101233226B (zh) 2005-06-22 2017-08-11 阿斯特利亚斯生物治疗股份公司 人胚胎干细胞的悬浮培养物

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0695351A1 (fr) * 1993-04-21 1996-02-07 University Of Edinburgh Isolation, selection et propagation de cellules souches d'animaux transgeniques
WO1996016162A1 (fr) * 1994-11-21 1996-05-30 National Jewish Center For Immunology And Respiratory Medicine Nouvelles populations de cellules embryonnaires et procedes d'isolation de ces populations

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5453366A (en) * 1990-07-26 1995-09-26 Sims; Michele M. Method of cloning bovine embryos

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0695351A1 (fr) * 1993-04-21 1996-02-07 University Of Edinburgh Isolation, selection et propagation de cellules souches d'animaux transgeniques
WO1996016162A1 (fr) * 1994-11-21 1996-05-30 National Jewish Center For Immunology And Respiratory Medicine Nouvelles populations de cellules embryonnaires et procedes d'isolation de ces populations

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
EVAN M J ET AL: "ESTABLISHMENT IN CULTURE OF PLURIPOTENTIAL CELLS FROM MOUSE EMBRYOS" NATURE, MACMILLAN JOURNALS LTD. LONDON, GB, vol. 292, 9 July 1981 (1981-07-09), pages 154-156, XP000941713 ISSN: 0028-0836 *
KELLER GORDON ET AL: "Hematopoietic commitment during embryonic stem cell differentiation in culture." MOLECULAR AND CELLULAR BIOLOGY, vol. 13, no. 1, 1993, pages 473-486, XP001104957 ISSN: 0270-7306 *
NICHOLS J ET AL: "DERIVATION OF GERMLINE COMPETENT EMBRYONIC STEM CELLS WITH A COMBINATION OF INTERLEUKIN-6 AND SOLUBLE INTERLEUKIN-6 RECEPTOR" EXPERIMENTAL CELL RESEARCH, SAN DIEGO, CA, US, vol. 215, no. 1, 1 November 1994 (1994-11-01), pages 237-239, XP002044063 ISSN: 0014-4827 *
PALACIOS R ET AL: "IN VITRO GENERATION OF HEMATOPOIETIC STEM CELLS FROM AN EMBRYONIC STEM CELL LINE" PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF USA, NATIONAL ACADEMY OF SCIENCE. WASHINGTON, US, vol. 92, August 1995 (1995-08), pages 7530-7534, XP000941683 ISSN: 0027-8424 *
See also references of WO9943785A1 *

Also Published As

Publication number Publication date
WO1999043785A1 (fr) 1999-09-02
JP2002504362A (ja) 2002-02-12
CA2320423A1 (fr) 1999-09-02
EP1056835A4 (fr) 2003-01-15
AU2879999A (en) 1999-09-15
AU757036B2 (en) 2003-01-30

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