Method of differentiation of morula or inner cell mass cells and method of making lineage-defective embryonic stem cells

• US 8,753,884 B2
• Filed: 05/16/2008
• Issued: 06/17/2014
• Est. Priority Date: 10/15/1999
• Status: Expired due to Fees

• Current Assignee: Advanced Cell Technology Incorporated (Astellas Pharma Incorporated)
• Original Assignee: Advanced Cell Technology Incorporated (Astellas Pharma Incorporated)
• Inventors: Cibelli, Jose, West, Michael D., Lanza, Robert
• Primary Examiner(s): Ton, Thaian N
• Application Number:
  US12/152,779
• Publication Number:
  US 20090104697A1
• Time in Patent Office: 2,223 Days
• Field of Search: 435/377, 435/366, 435/378, 435/379, 435/320.1
• US Class Current: 435/377
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 13/02 of urine or of the urinary tract, e.g. urine acidifiers
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 19/04 for non-specific disorders of the connective tissue
• CPC Class Codes: A61P 21/04 for myasthenia gravis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/14 for treating abnormal movements, e.g. chorea, dyskinesia
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/02 Non-specific cardiovascular stimulants, e.g. drugs for syncope, antihypotensives
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0606 Pluripotent embryonic cells, e.g. embryonic stem cells [ES] embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696
• CPC Class Codes: C12N 5/16 Animal cells

Abstract

An improved method of producing differentiated progenitor cells comprising obtaining inner cell mass cells from a blastocyst and inducing differentiation of the inner cell mass cells to produce differentiated progenitor cells. The differentiated progenitor cells may be transfected such that there is an addition, deletion or alteration of a desired gene. The differentiated progenitor cells are useful in cell therapy and as a I source of cells for the production of tissues and organs for transplantation. Also provided is a method of producing a lineage-defective human embryonic stem cell.

Claims

1. A method of producing differentiated cells, comprising:

i) obtaining morula cells or inner cell mass cells from a human blastocyst by mechanical and/or enzymatic means; and

ii) inducing the morula cells or inner cell mass cells to differentiate directly into differentiated cells, wherein the inducing step comprises a) addition of a differentiating agent directly to the morula cells or inner cell mass cells or b) placement of the morula cells or inner cell mass cells directly in a differentiation environment chosen from a flat adhesive environment, a microgravity environment and a 3D adhesive environment; and

iii) isolating the differentiated cells from ii), thereby producing differentiated cells.

2. The method according to claim 1, wherein said blastocyst is produced by in vitro fertilization.

3. The method according to claim 1, wherein said blastocyst is produced from a nuclear transfer unit.

4. The method according to claim 3, wherein a desired DNA is inserted, removed or modified in said nuclear transfer unit, thereby resulting in the production of a genetically altered differentiated cell.

5. The method according to claim 1, wherein said morula cells or inner cell mass cells are induced to differentiate in a flat adhesive environment.

6. The method according to claim 1, wherein said morula cells or inner cell mass cells are induced to differentiate in a 3D adhesive environment.

7. The method according to claim 1, wherein said morula cells or inner cell mass cells are induced to differentiate in a microgravity.

9. The method according to claim 1, wherein the differentiated cells are differentiated progenitor cells.

10. The method according to claim 9, wherein the differentiated progenitor cells give rise to mesoderm.

11. The method according to claim 9, wherein the differentiated progenitor cells give rise to ectoderm.

12. The method according to claim 9, wherein the differentiated progenitor cells give rise to endoderm.

8. A method of producing differentiated cells, comprising:

i) obtaining morula cells or inner cell mass cells from a human blastocyst by mechanical and/or enzymatic means; and

ii) inducing the morula cells or inner cell mass cells to differentiate directly into differentiated cells, wherein the inducing step comprises a) addition of a differentiating agent directly to the morula cells or inner cell mass cells or b) placement of the morula cells or inner cell mass cells directly in a differentiation environment wherein the morula or inner cell mass cells are induced to differentiate by generation of teratomas in immunodeficient mice; and

iii) isolating the differentiated cells from ii), thereby producing differentiated cells.

13. A method of producing differentiated cells, the method comprising:

i) obtaining morula cells or inner cell mass cells from a human blastocyst by mechanical and/or enzymatic means; and

ii) inducing the morula cells or inner cell mass cells to differentiate directly into differentiated cells, wherein the inducing step consists of a) addition of a differentiating agent directly to the morula cells or inner cell mass cells or b) placement of the morula cells or inner cell mass cells directly in a differentiation environment chosen from a flat adhesive environment, a microgravity environment, and a 3D adhesive environment, thereby producing

differentiated cells.

14. A method of producing differentiated cells, the method consisting of;

i) obtaining morula cells or inner cell mass cells from a human blastocyst by mechanical and/or enzymatic means; and

ii) inducing the morula cells or inner cell mass cells to differentiate directly into differentiated cells, wherein the inducing step consists of a) addition of a differentiating agent directly to the morula cells or inner cell mass cells or b) placement of the morula cells or inner cell mass cells directly in a differentiation environment chosen from a flat adhesive environment, a microgravity environment and a 3D adhesive environment; and

iii) isolating the differentiated cells from ii), thereby producing differentiated cells.

15. A method comprising:

i) obtaining morula cells or inner cell mass cells from a human blastocyst by mechanical and/or enzymatic means; and

ii) contacting the obtained morula cells or inner cell mass cells with a differentiating agent.