Structured nanocoatings for the stabilization of pluripotent stem cell media components

Patent metadata

Field	Value
Publication number	US-12465573-B2
Application number	US-201917045293-A
Country	US
Kind code	B2
Filing date	Apr 5, 2019
Priority date	Apr 6, 2018
Publication date	Nov 11, 2025
Grant date	Nov 11, 2025

Abstract

Disclosed are compositions and methods for pluripotent stem cell culture. Compositions include mineral coated microparticles having a core and a mineral coating, wherein the mineral coating includes fibroblast growth factor. Also disclosed are methods for pluripotent stem cell culture methods using mineral coated microparticles including fibroblast growth factor.

First claim

What is claimed is:

1 . A pluripotent stem cell culture method for maintaining stem cell pluripotency, the method comprising: contacting a pluripotent stem cell with a mineral coated microparticle, wherein the mineral coated microparticle comprises a core and a mineral coating that comprises fibroblast growth factor in an amount of fibroblast growth factor ranging from about 0.1 ng fibroblast growth factor per mg mineral coated microparticle to about 10,000 ng fibroblast growth factor per mg mineral coated microparticle, and wherein stem cell pluripotency is maintained when a minimum of 95% Oct4 + /Nanog + cell population is maintained at each cell passage.

2 . The method of claim 1 , wherein the fibroblast growth factor is selected from the group consisting of purified fibroblast growth factor, recombinant fibroblast growth factor, and combinations thereof.

3 . The method of claim 1 , wherein the fibroblast growth factor is basic fibroblast growth factor.

4 . The method of claim 1 , wherein the pluripotent stem cell is contacted with the mineral coated microparticle using a cell culture insert or the pluripotent stem cell is directly contacted with the mineral coated microparticle.

5 . The method of claim 1 , further comprising analyzing a cell for a marker selected from the group consisting of Oct4, Nanog, Sox2, SSEQ-4, TRA-1-60, TRA-1-81, and combinations thereof.

6 . The method of claim 3 , wherein the mineral coating comprises the basic fibroblast growth factor in an amount of about 1 μg per mg mineral coated microparticle.

7 . The pluripotent stem cell culture method of claim 1 , wherein the mineral coated microparticle is added at a concentration less than 0.5 mg/ml and the fibroblast growth factor is greater than 380 ng/ml.

8 . A pluripotent stem cell culture method of claim 1 , wherein the mineral coated microparticle is added at a concentration less than 0.5 mg/ml, and wherein the fibroblast growth factor ranges from greater than 760 ng fibroblast growth factor per mg mineral coated microparticle to less than or equal to 7,026 ng fibroblast growth factor per mg mineral coated microparticle.

Assignees

• Wisconsin Alumni Res Found

Inventors

• Murphy William L
• Khalil Andrew
• Xie Angela
• Johnson Hunter

Classifications

• C12N2533/90

  Substrates of biological origin, e.g. extracellular matrix, decellularised tissue · CPC title

• C12N2533/18

  Calcium salts, e.g. apatite, Mineral components from bones, teeth, shells · CPC title

• C12N2531/00

  Microcarriers · CPC title

• C12N2501/15

  Transforming growth factor beta (TGF-β) · CPC title

• C12N2501/115

  Basic fibroblast growth factor (bFGF, FGF-2) · CPC title