Remote monitoring of iPSC-derived neural crest cells

Dysfunctions of the human nervous system are a major health care problem. In order to understand and treat these dysfunctions in vitro research is performed on cells of the human nervous system, such as sensory neurons and neural crest cells. Neuronal cells used for in vitro research are often derived via directed differentiation of human-induced pluripotent stem cells (iPSCs), which can be a long (up to 30 days) and tedious process. In order to make sure the iPSCs differentiate into neuronal cells, it is very important to keep a close eye on your cells during this costly 30-day process. CytoSMART’s live-cell imaging microscopes can aid in this process.

Live-cell imaging to monitor your cells

Using the CytoSMART Lux2, for example, you can monitor the differentiation and proliferation of neuronal stem cells inside the incubator. Because all images are uploaded to the CytoSMART Cloud, you can follow the differentiation process without disturbing the iPSCs or even needing to go to the lab.

7-day neural differentiation method

However, even with monitoring, this long differentiation process is still prone to errors. Therefore, Anatomic has created a revolutionary method to differentiate human iPSCs into human sensory neurons through a neural crest intermediate state within just 7 days. Following one week of additional maturation, these sensory neurons have expression profiles similar to native human dorsal root ganglia with respect to key pain targets, begin firing action potentials, and possess sodium currents useful in studying pain. This rapid process allows you to scale out or scale up your research to the human nervous system dysfunctions.

Live-cell imaging of fast differentiated neural crest cells

CytoSMART and Anatomic have combined their strengths and used CytoSMART Lux2 to monitor the proliferation and function of neural crest cells, the intermediate state of the 7-day differentiation process. Neural crest cells are widely investigated in vitro to gain a better understanding of the formation of the nervous system but also of collective mesenchymal migration.

Time-lapse imaging of iPSC derived neural crest cell proliferation

Using Anatomic’s manufacturing platform, neural crest cells emerged from differentiating human iPSC cultures within just 4 days. In the movie below, harvested neural crest cell proliferation was monitored using the CytoSMART Lux2 for 3 days.

Time-lapse imaging of iPSC derived neural crest cell migration

The migration potential of the neural crest cells was investigated using the widely used scratch assay. A wound was created in the monolayer of neural crest cells using a pipette tip. The closure of the wound was monitored for 32 h at a 15 minute time interval in the video below.

Conclusion

In order to speed up the research into human nervous system dysfunctions, the 7-day differentiation method by Anatomic can be combined with live-cell imaging using the CytoSMART Lux2. In this way, the differentiation process is four times shorter than normal and by closely monitoring you can keep track of any irregularities in the differentiation process. More information about Anatomic and CytoSMART can be found below.

About Anatomic

Anatomic is helping to cure chronic pain conditions by delivering functional, consistent, and scalable stem cell-derived sensory neurons. Based on Anatomic’s breakthrough manufacturing platform, these Sensory Neurons enable the drug discovery industry to fail fast and accelerate the development of new, non-addictive pain killers. Anatomic is a privately held company based in Minneapolis.

About CytoSMART

CytoSMART is an innovator in kinetic live-cell imaging. Combining compact and fast imaging hardware with powerful image analysis algorithms supported by cloud computing. Automation in time-lapse microscopy and image-based cell counting to generate high-quality and robust data. Our team of engineers continues to develop and optimize the image analysis and data storage capacities linked to our systems, making sure that data sets are easily processed, stored, and kept securely in an online environment. We want to enable research that builds on large data sets and is conveniently communicated.