Methods for Accelerated and Enhanced Cardiac Differentiation of IPS Cells by Electrical Stimulation

Patent metadata

Field	Value
Publication number	US-2016201035-A1
Application number	US-201614995997-A
Country	US
Kind code	A1
Filing date	Jan 14, 2016
Priority date	Jan 14, 2015
Publication date	Jul 14, 2016
Grant date	—

Abstract

Methods of generating cardiomyocytes from induced pluripotent stem cells (IPSCs) are provided. More specifically, the present disclosure relates to methods of generating cardiomyocytes from iPSCs using electrical stimulation. In some aspects, uses of such cells for therapeutics and in methods of treatment are provided.

First claim

What is claimed:

1 . A method for the accelerated and enhanced generation of autologous cardiac cells, the method comprising: applying electrical stimulation to induced pluripotent stem cells for a time period to induce cardiomyocyte differentiation of the induced pluripotent stem cells to form cardiomyocytes without the use of exogenous growth factors, wherein the exogenous growth factors are selected from the group consisting of bone morphogenetic proteins, wingless/INT proteins, fibroblastic growth factors, vascular endothelial growth factor, activin A and ascorbic acid/vitamin C.

2 . The method of claim 1 , wherein induced pluripotent stem cells comprise embryoid bodies.

3 . The method of claim 1 , wherein the electrical stimulation is a biomimetic electrical stimulation.

4 . The method of claim 1 , wherein the electrical stimulation is applied to the induced pluripotent stern cells at an electric field strength of from about 1 V/1.8 cm to about 1.5 V/1.8 cm.

5 . The method of claim 4 , wherein the electrical stimulation is applied to the induced pluripotent stem cells using a biphasic square pulse.

6 . The method of claim 5 , wherein the biphasic square pulse is applied to the induced pluripotent stem cells over about a 5 ms period.

7 . The method of claim 6 , wherein the electrical stimulation is applied at about a 5 Hertz frequency.

8 . A method of treating a patient suffering from cardiac tissue deficiency, damage, or loss, the method comprising:
applying electrical stimulation to induced pluripotent stem cells for a time period to induce cardiomyocyte differentiation of the induced pluripotent stem cells to form cardiomyocytes without the use of exogenous growth factors, wherein the exogenous growth factors selected from the group consisting of bone morphogenetic proteins, wingless/INT proteins, fibroblastic growth factors, vascular endothelial growth factor, activin A and ascorbic acid/vitamin C; and delivering an effective amount of one or more of the formed cardiomyocytes into a cardiovascular system component of the patient.

9 . The method according to claim 8 , wherein the cardiomyocytes are autologous cardiomyocytes.

10 . The method of claim 9 , wherein the electrical stimulation is a biomimetic electrical stimulation.

11 . The method of claim 9 , wherein the electrical stimulation is applied to the induced pluripotent stem cells at an electric field strength of from about 1 V/1.8 cm to about 1.5 V/1.8 cm.

12 . The method of claim 11 , wherein the electrical stimulation is applied to the induced pluripotent stem cells using a biphasic square pulse.

13 . The method of claim 12 , wherein the biphasic square pulse is applied to the induced pluripotent stem cells over about a 5 ins period.

14 . The method of claim 13 , wherein the electrical stimulation is applied at about a 5 Hertz frequency.

15 . The method of claim 8 , wherein delivering comprises injection or implantation.

16 . The method according to claim 15 , wherein delivering comprises transplantation and the cardiomyocytes are delivered as a tissue.

17 . The method according to claim 8 , wherein the cardiovascular system component is the heart.

18 . A method for the accelerated and enhanced generation of autologous cardiac cells, the method comprising:
seeding induced pluripotent stem cells on a substrate; cultivating the seeded induced pluripotent stem cells without exogenous growth factors, wherein the exogenous growth factors selected from the group consisting of bone morphogenetic proteins, wingless/INT proteins, fibroblastic growth factors, vascular endothelial growth factor, activin A and ascorbic acid/vitamin C; and applying electrical stimulation to the seeded induced pluripotent stem cells for a time period to induce cardiomyocyte differentiation of the induced pluripotent stern cells to form cardiomyocytes.

19 . The method of claim 18 , wherein the electrical stimulation is a biomimetic electrical stimulation.

20 . The method of claim 18 , wherein the substrate comprises an extracellular matrix protein.

Assignees

• Univ Cincinnati

Inventors

• Wang yi-gang
• Kim Ha Won
• Ma Ruilian

Classifications

• C12N2506/45

  from artificially induced pluripotent stem cells · CPC title

• C12N2529/00

  Culture process characterised by the use of electromagnetic stimulation · CPC title

• C12N5/0657 Primary

  Cardiomyocytes; Heart cells · CPC title

• A61K35/34

  Muscles; Smooth muscle cells; Heart; Cardiac stem cells; Myoblasts; Myocytes; Cardiomyocytes (vascular smooth muscle A61K35/44) · CPC title

• C12N5/0696

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