Method for expansion of stem cells

• US 8,895,299 B2
• Filed: 06/25/2010
• Issued: 11/25/2014
• Est. Priority Date: 02/15/2005
• Status: Active Grant

• Current Assignee: Xon Cells, Inc (Precigen, Inc.)
• Original Assignee: Xon Cells, Inc (Precigen, Inc.)
• Inventors: Riordan, Neil H.
• Primary Examiner(s): Kim, Taeyoon
• Application Number:
  US12/823,960
• Publication Number:
  US 20100261152A1
• Time in Patent Office: 1,614 Days
• Field of Search: 424/93.7, 435/325, 435/404
• US Class Current: 435/325
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: A61K 35/44 Vessels; Vascular smooth muscle cells; Endothelial cells; Endothelial progenitor cells
• CPC Class Codes: A61K 35/50 Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells
• CPC Class Codes: A61K 35/51 Umbilical cord; Umbilical cord blood; Umbilical stem cells
• CPC Class Codes: A61K 38/1825 Fibroblast growth factor [FGF]
• CPC Class Codes: A61K 8/982 Reproductive organs; Embryos, Eggs
• CPC Class Codes: A61Q 19/08 Anti-ageing preparations
• CPC Class Codes: A61Q 7/00 Preparations for affecting hair growth
• CPC Class Codes: C12N 2502/02 embryonic cells
• CPC Class Codes: C12N 5/0018 Culture media for cell or tissue culture media for specific animal cell type C12N5/06

Abstract

A method of increasing the growth of stem cells by mixing the stem cells with a growth medium that has been conditioned by an incubation with placental tissue. The method increases the expansion of the stem cell population.

Claims

1. A method for the expansion or growth of stem cells without substantially inducing differentiation, the method comprising:

collecting cord blood from a human umbilical cord;

isolating CD34+ cells from the cord blood;

producing live placenta conditioned medium (LPCM) by incubating whole human placenta in a Dulbecco'"'"'s modified Eagle medium (DMEM) in a sterile container, wherein the whole placenta is submerged;

isolating the LPCM; and

contacting at least one CD34+ cell with a supplemented DMEM comprising the LPCM and a cytokine cocktail comprising 20 ng/ml IL-3, 250 ng/ml IL-6, 10 ng/ml stem cell factor, 250 ng/ml thrombopoietin, and 100 ng/ml Flt-3L, wherein the concentration of the LPCM in the supplemented DMEM is 50%.

2. The method of claim 1, wherein isolating CD34+ cells comprises a method selected from the group consisting of fluorescence activated cell sorting, magnet activated cell sorting, counterflow centrifugal elutriation, equilibrium density centrifugation, velocity sedimentation at unit gravity, and T lymphocyte depletion.

3. The method of claim 1, wherein incubating whole human placenta in a DMEM comprises a period of at least 24 hours.

4. The method of claim 1, further comprising decontaminating the isolated LPCM by a method selected from the group consisting of filtration, UV irradiation, X-ray sterilization, ozonation, and hyperthermia.

5. The method of claim 1, wherein contacting at least one CD34+ cell with a supplemented DMEM comprises a period of at least 24 hours.

6. The method of claim 1, wherein the supplemented DMEM comprises a growth factor selected from the group consisting of, IL-1, IL-7, G-CSF, GM-CSF, Epo, FGF-1, FGF-2, FGF-4, FGF-20, IGF, EGF, NGF, LIF, PDGF, bone morphogenic proteins (BMP), activin-A, VEGF, and forskolin.