Gynogenetic or androgenetic production of pluripotent cells and cell lines, and use thereof to produce differentiated cells and tissues

• US 7,951,591 B2
• Filed: 02/27/2003
• Issued: 05/31/2011
• Est. Priority Date: 10/28/1999
• Status: Expired due to Fees

• Current Assignee: University Of Massachusetts
• Original Assignee: Advanced Cell Technology Incorporated (Astellas Pharma Incorporated)
• Inventors: Robl, James M., Cibelli, Jose, Burnside, Amy
• Primary Examiner(s): Crouch; Deborah
• Assistant Examiner(s): NOBLE, MARCIA STEPHENS
• Application Number:
  US10/374,512
• Publication Number:
  US 20040014206A1
• Time in Patent Office: 3,015 Days
• Field of Search: 435/325, 435/366, 435/455, 435/377, 435/373, 435/384
• 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: A61K 35/545 Embryonic stem cells; Pluripotent stem cells; Induced pluripotent stem cells; Uncharacterised stem cells
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 15/873 Techniques for producing new embryos, e.g. nuclear transfer, manipulation of totipotent cells or production of chimeric embryos
• CPC Class Codes: C12N 2500/14 Calcium; Ca chelators; Calcitonin
• CPC Class Codes: C12N 2501/999 Small molecules not provided for elsewhere
• CPC Class Codes: C12N 2502/13 connective tissue cells; generic mesenchyme cells, e.g. so-called "embryonic fibroblasts"
• CPC Class Codes: C12N 2506/04 from germ cells
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 2517/10 Conditioning of cells for in vitro fecondation or nuclear transfer
• 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 5/0696 Artificially induced pluripotent stem cells, e.g. iPS
• CPC Class Codes: G01N 2333/475 Assays involving growth factors
• CPC Class Codes: G01N 33/5008 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 33/502 for testing non-proliferative effects
• CPC Class Codes: G01N 33/5044 involving specific cell types
• CPC Class Codes: G01N 33/5073 Stem cells
• CPC Class Codes: G01N 33/5088 of vertebrates

Abstract

Methods for obtaining pluripotent (embryonic stem) cells from parthenogenetic embryos, especially primates, are provided. These cells are useful for producing differentiated cells, tissues and organs, especially human and non-human primate cells, tissues and organs.

Claims

1. A method for producing a pluripotent cell comprising:

(a) obtaining a primate oocyte in metaphase II, which optionally may be genetically modified;

(b) activating said oocyte by a method selected from the group consisting of

(i) culturing the oocyte under conditions that do not result in second polar body extrusion;

(ii) culturing the oocyte in the presence of an agent that inhibits polar body extrusion, and

(iii) culturing the oocyte under conditions that prevent the initial cleavage of the activated oocyte;

(c) culturing said activated oocyte to produce a gynogenetic embryo comprising a discernible trophectoderm and an inner cell mass;

(d) isolating said inner cell mass or cells therefrom and transferring said inner cell mass or cells therefrom to an in vitro media that inhibits differentiation; and

(e) culturing said inner cell mass or cells therefrom to maintain said cells in an undifferentiated pluripotent state.

2. The method of claim 1, further comprising differentiating in vitro said cultured cells of (e).

3. The method of claim 1, further comprising implanting one or more of said cultured cells of (e) into an immunocompromised non-human animal, thereby forming a teratoma in said non-human animal.

4. The method of claim 1, wherein the primate oocyte is a monkey or human oocyte.

5. The method of claim 4, wherein the primate oocyte is a human oocyte.

6. The method of claim 1, wherein the primate oocyte is a non-human primate oocyte.

7. The method of claim 1, wherein said agent that inhibits polar body extrusion is DMAP.

8. The method of claim 1, wherein said activation method in (b) (iii) comprises culturing the oocyte in the presence of a compound that inhibits microfilament formation or protein production thereby preventing the initial cleavage of the activated oocyte.

9. The method of claim 8, wherein said compound is cycloheximide or cytochalasin B.

10. The method of claim 1, wherein the primate oocyte of (a) is genetically modified.