Human IPSC-derived vascular-related and hematopoetic cells for therapies and toxicology/drug screenings

Patent metadata

Field	Value
Publication number	US-10385313-B2
Application number	US-201415026313-A
Country	US
Kind code	B2
Filing date	Oct 1, 2014
Priority date	Oct 1, 2013
Publication date	Aug 20, 2019
Grant date	Aug 20, 2019

Abstract

Described herein are cells, cell culture methods, and cell culture media compositions useful for producing and maintaining iPSC-derived cell lines that are of higher purity and maintain cell type integrity better than current iPSC-derived cell lines. Also disclosed are methods of using the described cells and media, such as therapeutic methods of use for the described cells. The described cells include iPSC-derived mesodermal precursor cells (MPC), which itself may differentiate into at least four different cell types. When cultured under appropriate conditions, the mesodermal precursor cells can be used to produce hematopoietic stem cells (HSC), mesenchymal stem cells (MSC), smooth muscle cells (SMC), or unlimited functional endothelial cells (UFEC). One characteristic that makes the described cells desirable is that they can be maintained in culture for a number of days, or passages, without changing phenotype through differentiation.

First claim

What is claimed is:

1. A cell culture medium consisting essentially of:
Iscove's modified Dulbecco's medium (IMDM),
Ham's F-12 Nutrient Mix, with L-alanyl-L-glutamine additive,
Albumin,
a-monothioglycerol,
protein-free hybridoma mixture II,
L-ascorbic acid 2-phosphate,
L-alanyl-L-glutamine,
Antibiotic,
insulin-transferrin-selenium-ethanolamine supplement,
bone morphogenic protein 4,
vascular endothelial growth factor, and
basic fibroblast growth factor.

2. The cell culture medium of claim 1 , further comprising cholesterol lipids.

3. The cell culture medium of claim 1 , wherein the antibiotic is selected form the group consisting of penicillin, streptomycin and a mixture of penicillin and streptomycin.

4. The cell culture medium of claim 1 , wherein the concentration of albumin, is about 5 mg/ml.

5. The cell culture medium of claim 1 , wherein the concentration of monothioglycerol is from about 350 to about 450 μM.

6. The cell culture medium of claim 1 , wherein the concentration of L-ascorbic acid 2-phosphate is about 50 μg/ml.

7. The cell culture medium of claim 1 , wherein the concentration of L-alanyl-L-glutamine is about 1 mM to about 2 mM.

8. The cell culture medium of claim 1 , wherein the concentration of bone morphogenic protein 4 is about 10 ng/ml.

9. The cell culture medium of claim 1 , wherein the concentration of vascular endothelial growth factor is about 10 ng/ml.

10. The cell culture medium of claim 1 , wherein the concentration of basic fibroblast growth factor is about 10 ng/ml.

Assignees

• Us Health
• The Usa As Represented By The Sec Dep Of Health And Human Services

Inventors

• Boehm Manfred
• Chen Guibin
• Rao Mahendra
• Larochelle Andre

Classifications

• C12N5/0696

  Artificially induced pluripotent stem cells, e.g. iPS · CPC title

• C12N2501/26

  Flt-3 ligand (CD135L, flk-2 ligand) · CPC title

• C12N5/0018 Primary

  Culture media for cell or tissue culture (media for specific animal cell type C12N5/06) · CPC title

• C12N2500/44

  Thiols, e.g. mercaptoethanol · CPC title

• C12N2500/30

  Organic components · CPC title