Multipotent prenatal stem cells

• US 9,700,585 B2
• Filed: 09/26/2013
• Issued: 07/11/2017
• Est. Priority Date: 09/26/2013
• Status: Active Grant

• Current Assignee: Prime Merger Sub LLC (Alight, Inc.)
• Original Assignee: NuTech Medical Incorporated (Organogenesis Holdings, Inc.)
• Inventors: Vines, Jeremy B., Walthall, Jr., Howard P.
• Primary Examiner(s): FOX, ALLISON M
• Application Number:
  US14/038,274
• Publication Number:
  US 20150216911A1
• Time in Patent Office: 1,384 Days
• Field of Search: None
• CPC Class Codes: A61K 35/50 Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells
• CPC Class Codes: C12N 5/0605 Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly
• CPC Class Codes: C12N 5/0668 Mesenchymal stem cells from other natural sources

Abstract

The present invention relates to the isolation, propagation and use of prenatal stem cells. The cells are CD105 negative. In certain embodiments the cells are also SSEA-4 positive or negative, and c-kit negative. The cells may be isolated from a prenatal sample, for instance third trimester amniotic fluid or placental membrane. The cells may be combined with a biologically compatible solution or a biologically compatible matrix and utilized to treat a human subject. The invention also contemplated enriched populations of prenatal stem cells. The enriched cell population may be propagated in the appropriate culture medium and the resulting progeny cells utilized therapeutic applications.

Claims

1. A method of harvesting human prenatal stem cells comprising:

providing a human prenatal sample;

collecting cells from the sample;

selecting CD105 negative cells from the sample, and

selecting from the CD 105 negative cells, cells that are CD44 positive, CD90 positive or both CD44 positive and CD90 positive thereby obtaining CD105 negative cells that are CD44 positive, CD90 positive or both CD44 positive and CD90 positive.

2. The method of claim 1, wherein the prenatal sample comprises prenatal fluid.

3. The method of claim 2, wherein the prenatal fluid comprises third trimester amniotic fluid.

4. The method of claim 1, wherein the prenatal sample comprises prenatal membrane.

5. The method of claim 4, wherein the prenatal membrane comprises placental membrane.

6. The method of claim 1, wherein the selecting of the CD105 negative cells comprises the steps of:

incubating a plurality of cells from the human prenatal sample with a first antibody reactive to CD105; and

selecting from the plurality of cells a group of cells that are not reactive with the first antibody.

7. The method of claim 6, wherein the selecting from the plurality of cells a group of cells that are not reactive with the first antibody is accomplished by an immunoselection technique.

8. The method of claim 7, where the immunoselection technique is selected from the group consisting of flow cytometry, activated cells sorting and immune-magnetic selection.

9. The method of claim 1, further comprising selecting c-kit negative cells from the CD105 negative cells.

10. A method of harvesting human prenatal stem cells comprising:

providing a human prenatal sample;

collecting cells from the sample;

selecting CD105 negative cells from the sample, wherein the CD105 negative cells are CD44 positive or both CD44 positive and CD90 positive, wherein the selecting of the CD105 negative cells comprises the steps of;

incubating a plurality of cells from the human prenatal sample with a first antibody reactive to CD105; and

selecting from the plurality of cells a group of cells that are not reactive with the first antibody

incubating the group of cells that are not reactive with the first antibody with a second antibody that is reactive to CD44; and

selecting from the group of cells that are not reactive with the first antibody a second group of cell that are reactive with the second antibody.

11. A method of proliferating a population of human prenatal stem cells comprising:

providing a human prenatal sample including CD105 negative cells and CD105 positive cells;

isolating the CD105 negative cells from the CD105 positive cells, the isolated CD105 negative cells having a greater proliferation rate than the isolated CD105 positive cells;

selecting from the isolated CD 105 negative cells, cells that are CD44 positive, CD90 positive or both CD44 positive and CD90 positive thereby obtaining CD105 negative cells that are CD44 positive, CD90 positive or both CD44 positive and CD90 positive; and

thereafter,

proliferating the CD105 negative cells that are CD44 positive, CD90 positive or both CD44 positive and CD90 positive.

12. A method of proliferating a population of human prenatal stem cells comprising:

providing a human prenatal sample including CD105 negative cells and CD105 positive cells;

isolating the CD105 negative cells from the CD105 positive cells, the isolated CD105 negative cells having a greater proliferation rate than the isolated CD105 positive cells and wherein the isolated CD105 negative cells include SSEA4 negative cells and SSEA4 positive cells;

proliferating the isolated CD105 negative cells;

isolating from the isolated CD105 negative cells the SSEA4 negative cells; and

proliferating the SSEA4 negative cells, the SSEA4 negative cells having a greater proliferation rate than the SSEA4 positive cells.