Enhanced throughput mineral coatings for optimization of stem cell behavior

Patent metadata

Field	Value
Publication number	US-10745739-B2
Application number	US-201815918357-A
Country	US
Kind code	B2
Filing date	Mar 12, 2018
Priority date	Feb 19, 2013
Publication date	Aug 18, 2020
Grant date	Aug 18, 2020

Abstract

Disclosed are methods for cell transfection and regulating cellular behavior. More particularly, the present disclosure relates to methods of non-viral cell transfection and regulating cellular behavior using mineral coatings that allow for the enhanced transfection of cells. The mineral coatings bind polynucleotides and provide a source of calcium and phosphate ions to enhance transfection. The present disclosure also provides a high throughput platform for screening non-viral transfection of cells. The methods of the present disclosure also provide an advantageous polynucleotide delivery platform because the mineral coatings may be deposited on various medical device materials after being specifically developed using the high throughput screening platform.

First claim

What is claimed is:

1. A non-viral transfection system comprising:
a substrate that comprises:
a mineral coating, wherein the mineral coating comprises a calcium to phosphate ratio of from about 2.5:1 to about 1:1;
a polynucleotide-lipid complex bound to the mineral coating; and
one or more cells.

2. The non-viral transfection system of claim 1 , wherein the substrate further comprises a poly(α-hydroxy ester).

3. The non-viral transfection system of claim 2 , wherein the poly(α-hydroxy ester) is selected from the group consisting of poly(L-lactide), poly(lactide-co-glycolide), poly(ε-caprolactone), and combinations thereof.

4. The non-viral transfection system of claim 1 , wherein the substrate is selected from the group consisting of a film, a plate, a dish, a coverslip, and a slide.

5. The non-viral transfection system of claim 1 , wherein the mineral coating comprises a morphology selected from the group consisting of a spherulitic microstructure, a plate-like microstructure, a net-like microstructure, and combinations thereof.

6. The non-viral transfection system of claim 5 , wherein the spherulitic microstructure comprises an average spherulite diameter of from about 2 μm to about 4 μm.

7. The non-viral transfection system of claim 1 , wherein the substrate comprises more than one well or more than one chamber.

8. The non-viral transfection system of claim 1 , wherein the cell is selected from the group consisting of a pluripotent stem cell, a mesenchymal stem cell, and an umbilical vein endothelial cell.

9. The non-viral transfection system of claim 1 , wherein the substrate comprises a three-dimensional scaffold.

10. A non-viral transfection system, comprising:
a substrate, wherein the substrate is three-dimensional poly(α-hydroxy ester) scaffold,
a mineral coating on the surface of the substrate, wherein the mineral coating comprises a morphology selected from the group consisting of a spherulitic microstructure, a plate-like microstructure, a net-like microstructure, and combinations thereof;
a polynucleotide-lipid complex bound to the mineral coating; and
one or more cells, wherein the cell is selected from the group consisting of a pluripotent stem cell, a mesenchymal stem cell, and an umbilical vein endothelial cell.

Assignees

• Wisconsin Alumni Res Found

Inventors

• Murphy William L
• Choi Siyoung
• Yu Xiaohua

Classifications

• C12N15/87 Primary

  Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation · CPC title

• C12Q1/68 Primary

  involving nucleic acids · CPC title