Scalable, xeno-free cardiomyocyte differentiation protocol
Scalable, xeno-free cardiomyocyte differentiation protocol
A serum-free protocol and a serum-containing protocol to generate human cardiomyocytes from control or patient-derived iPSCs. The use of cheap, defined chemicals at a low dose eliminates the need for costly recombinant factors, substantially reducing the otherwise high input cost of producing human cardiomyocytes.
New York, NY, United States
Overview Comments Tagged publications
Background 

Given the prevalence of cardiac diseases, there is substantial demand for new therapeutics and improved methods for screening drug toxicity. Generating cardiomyocytes from human induced pluripotent stem cells (hiPSCs) offers a unique opportunity to study cardiac disease. However, the cost of the recombinant factors used and the requirement for xeno-free and feeder-free production in a pharmaceutical setting limit large scale production of human cardiomyocytes. This technology offers a serum-free protocol and a serum-containing protocol for generating cardiomyocytes from hiPSCs. Further, this technology offers an effective means for examining the excitation-contraction coupling of the human cardiomyocytes with a dual optical fluorescent reporter system that monitors calcium and voltage levels, to ensure cells derived from patients and healthy controls maintain their native characteristics. Together, this protocol enables healthy, native-like cardiomyocytes to be derived on a large scale suitable for screening therapeutics and drug toxicity tests.

Optimized protocols for low cost, efficient differentiation of hiPSCs into cardiomyocytes

This technology includes a serum-free protocol and a serum-containing protocol to generate human cardiomyocytes from control or patient-derived iPSCs. The use of cheap, defined chemicals at a low dose eliminates the need for costly recombinant factors, substantially reducing the otherwise high input cost of producing human cardiomyocytes. Analysis of in-vitro cultures demonstrates that this serum-free protocol offers higher yield than current methods. Further, this technology includes dual optical fluorescent reporters that are administered through lentiviral infection to record calcium and voltage simultaneously, allowing the cardiomyocytes to be characterized and monitored during extended time-course experiments, as well as during repeat experiments using the same cells. The cost-effectiveness and absence of animal derived products make this technology an important advance in generating human cardiomyocytes from iPSCs for use in screens for therapeutics and drug toxicity.

This technology has been tested in-vitro using both control and patient-derived cells. Cardiomyocytes derived from hiPSCs from patients with Timothy syndrome have been used and were determined to maintain the diseased phenotype.

Lead Inventor:

Masayuki Yazawa, Ph.D.

Applications:

  • Differentiates control and patient derived hiPSCs into cardiomyocytes
  • Large-scale production of cardiomyocytes
  • Therapeutic and drug-toxicity screens
  • Extended physiological experiments measuring excitation-contraction 
  • Cardiac tissue engineering 
  • Cardiac cell based therapies

Advantages:

  • Eliminates need for animal-derived products in producing cardiomyocytes
  • Utilizes inexpensive, defined chemicals at low doses 
  • Fluorescent reporter offers method to monitor calcium and voltage levels 
  • Protocols are applicable to control and patient-derived cells
  • Absolute yield of cardiomyocytes with a serum-free protocol is higher than other commonly used protocols
  • Feeder-free monolayer culture method increases convenience of maintaining the cells 

Patent Information:

Patent Pending

Tech Ventures Reference: IR CU15304

Related Publications:

  • [Song L, Awari D, Han E, Uche-Anya E, Park S, Yabe Y, Chung W, Yazawa M. «Dual optical recordings for action potentials and calcium handling in induced pluripotent stem cell models of cardiac arrhythmias using genetically encoded fluorescent indicators” Stem Cells Transl Med. 2015 May; 4(5): 468-475.] (http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4414223/ )
No comments.
No tagged publications.

Similar innovations

Back
or
Back
Back
or
Back
Back
or
Back
Back
or
Back
  • Rapid in vitro analysis of force strain in cell signaling pathways for use in tissue engineering
  • Facile method allows direct observation of effect of force with much higher throughput than other methods. Other methods to apply force to proteins, such as AFM or optical tweezers, are limited by the difficulty of setting up experiments and the time to complete an analysis. Additionally, these instruments are very expensive and difficult to operate and maintain. This method allows for multiple measurements to be conducted at the same time, greatly increasing the rate of experiments that can be conducted by a single researcher.
  • Read more
  • Methods
  • New York, NY, United States
  • Stanford University
← Previous Next → 1 2 3 4 Last
Showing 1-4 from 231

Send Us a Message