Directed Differentiation of Pluripotent Stem Cells

Sponsored by: Thermo Fisher Scientific
Session ends: May 22nd, 2015, 3:00pm MST
Answers by: Dr. Mohan C Vemuri , Thermo Fisher Scientific, Inc.


The promise of human pluripotent stem cells will be realized only when these cells are successfully coaxed into different cell types found in the human body, through the process of directed differentiation. This is critical to getting the desired cell types and numbers needed for drug screening, translational cell therapy and regenerative medicine applications. Most of the existing methods of differentiation are suboptimal, involving laborious mechanical and manual steps leading to issues of reproducibility and reduced efficiency in downstream processing of functionally mature lineages. The complex developmental process of differentiation and the challenges associated need to be efficiently deciphered in order to successfully direct the hPSC differentiation to target cell types.

During this Ask the Experts session, we will be discussing the challenges associated with hPSC differentiation to neural, cardiac, and definitive endoderm lineages, how Thermo Fisher Scientific can address how these processes can be efficiently simplified with tools and cGMP cell culture media systems for robust, efficient and scalable differentiation of these two critical cell lineages. Use of these reagent systems will enable researchers to precisely control and direct the differentiation to terminal lineages in a relatively easy manner, and speedily with high efficiency.

This Ask the Expert session is sponsored by Thermo Fisher Scientific and hosted by Dr. Mohan C Vemuri. Dr. Vemuri is the Director of Research and Development for Cell Biology at Thermo Fisher Scientific.

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Please take advantage of the opportunity to ask our expert a question and participate in a lively discussion on directed differentiation of human pluripotent stem cells.

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Questions & Answers

Why are many published differentiation protocols are not reproducible?

Many of these methods are suboptimal and a high percentage of methods even if published in highly reputed journals can not be reproduced. Variations come from the cell lines used, quality of the pluripotency state to begin with, use of undefined components in media and matrix used , and operator variations ( say a novice […]» Read More

We are looking for the simplest most successful method for differentiation cardiomyocytes. I have investigated 4 methods, we are most familiar with embyoid bodies but I have read that guided differentiation has the best efficiency. What do you recommend? Do you have a protocol that you have seen success with?

Cardiomyocyte differentiation through embryoid body methods is quite variable in addition to the interline variation from iPSCs. Thermofisher has developed a monolayer method of cardiac differentiation that bypasses EB formation, and one can achieve efficient differentiation with cardiomyocyte differentiation kit ( for example >90% cardiomyocytes using H9 line). The kit, supporting materials and detailed protocols […]» Read More

What is your preferred method for confirming differentiation?

Along the path of differentiation, ensuring early, middle and late stage specific marker expression for different lineages and finally testing the cells for their physiological function – for example, if the cells are being differentiated to dopaminergic neurons, analyzing the dopamine release in these cultures and finally testing the cells in in vivo animal transplants […]» Read More

I am currently using monolayer culture for differentiation of blood cells. I am looking at embryoid bodies and was wondering if there is a benefit to this method and also challenges with the method.

Which cell types are you starting with as monolayer is an important consideration for differentiation. If pluripotent stem cells (PSC)are started as monolayer and driving the differentiation towards Hematopoietic stem cells (HSC), starting as adherent monolayer of PSC and harvesting HSC as suspension culture in the same dish is an ideal set up. In contrast, […]» Read More