Stem cell research
Stem cell culture
monitoring using
non-invasive imaging
Stem cell sources such as mesenchymal stem cells, induced pluripotent stem cells and neural stem cells, are currently under investigation to be used as therapeutic agents.
Culturing stem cells is not straightforward, but their potential holds great promise in combating pathologies that are either poorly treatable or not treatable at all.
Stem cell culturing is a long-term process. From isolation to expansion to harvesting, the process can take weeks. Many cell labs have their own optimized protocols, which are meant to maintain stemness and stem cell proliferation. However, batch-to-batch differences can have a significant impact on the quality of the final product.
Non-invasive imaging and time-lapse imaging can provide straightforward solutions in both monitoring and optimization of stem cell cultures. Imaging modalities can provide researchers insight and control into their stem cell culture.
Image stem cells from within hypoxia chambers
Hands-off microscopy using cloud connection, so there is no need to take the cells out of the incubator.
Mesenchymal, hematopoietic, neural and embryonic stem cells are supplied with 1-8% oxygen in their physiological niche. This is a significant difference to the 21% that is usually maintained in incubators. Hypoxia chambers can help to ensure desirable stem cell viability, yield and stemness. Imaging cell cultures using a remote connection via cloud computing e.g. with the Lux2 can help to prevent environmental shock when acquiring microscopic data.
Multi-well analysis of proliferation
Automated time-lapse imaging and confluency detection
For high throughput assays and cell line optimization it is a great benefit to obtain kinetic information. The CytoSMART Omni is able to automatically create time-lapse videos of whole wells in multi-well plates. Cell culture growth rates, morphology changes and apoptotic events can be visualized and subsequently analyzed using automated image analysis.
The CytoSMART Omni is ideal to perform kinetic assays to map cell proliferation, which we use as a tool to understand drug resistance in pancreatic cancer cell lines.
Dr. Martin Sprick | Senior Group Leader
Heidelberg Institute for Stem Cell Technology and Experimental Medicine
Yield optimization by automated monitoring
Supported by automated confluency detection
The longevity of the culture process of stem cells requires patience and careful monitoring. Time-lapse imaging can help in keeping track of the culture confluence and any changes that occur in morphology. Especially for iPSCs it is important that the colonies are not over-growing which leads to unstable stem cells. The CytoSMART Lux2 Duo Kit can help in keeping track of cell cultures from within the incubator and employs automated confluency detection to monitor cultures closely and unambiguously. The CytoSMART Omni can scan full flask surfaces such as T-175 or HYPERflasks and can be used to inspect culture homogeneity.
Label-free imaging of live cells
Stem cells have distinct morphological characteristics and growth patterns. Being able to explore and quantify these can be greatly beneficial for culture optimization and assay development. Clonogenic assays and wound healing assays are standard assays that benefit from whole-well analysis. Alternatively for comparative single cell tracking we recommend the CytoSMART Lux2 Duo Kit.
Related articles
Are you looking for more information on stem cell types, their isolation, culturing methods and applications? Read our blog articles on induced pluripotent stem cells, neural stem cells and mesenchymal stem cells from the umbilical cord and bone marrow.
Videos
Appnotes
Cell Motility Video Monitoring - Lux2
Cell Cytoxicity Assay to Analyze Drug Response
Cell Culture Monitoring - Lux2
Cell Motility Video Monitoring - Lux2
Movement of cells plays a critical role in the development of cancer. Analyzing the motility of cells in vitro is therefor important for many cancer researchers.
Live-cell imaging, and in particular label-free live-cell imaging, is well suited to capture dynamic processes in cell culture without potential side-effects of markers or dyes on the cells. In the text below we will discuss the suitability of the CytoSMARTTM
Lux for several assays relevant for cancer research
We will discuss
- Introduction to the Lux cell monitoring system
- Tube formation assays
- 2D migration assays
- 3D invasion assays
Cell Cytoxicity Assay to Analyze Drug Response
The effect of drugs and drug candidates on the viability of cells in culture can be determined using cell counting, live/dead assays and metabolic assays. However, these assays are often end-point measurements. Alternatively, cells can be monitored using bright-field microscopy, by creating time-lapse videos for a culture period of multiple days. To study the lasting effect of the drug candidate.
In this study the cytotoxic effect of a chemotherapy drug was investigated for a range of concentrations. The effect on cell viability between drug concentrations was compared by analyzing confluency measurements obtained using automated live-cell imaging. The entire experiment was performed inside a CO2-incubator, ensuring optimal culturing conditions and cells were imaged every hour for a period of 3 days.
Cell Culture Monitoring - Lux2
Setting up cell cultures is easy enough. However monitoring your cultures and optimization is time-consuming and cumbersome. Waiting for the ideal confluency, quickly studying effects of various media means taking your cells in and out of the incubator more often than you would like.
Visualizing cell cultures from inside an incubator using a compact microscope that facilitates live cell imaging can overcome these issues. While live cell imaging has been restricted to costly, high-end devices, the CytoSMART Lux2 offers an affordable and easy-to-use alternative for virtually any lab. The CytoSMART Lux2 can be set up in minutes, enabling untrained users to quickly perform their own time-lapse recordings.
Images and videos can be easily accessed and retrieved from the CytoSMART cloud portal. Advanced functions, such as reporting of cell confluency, cell migration analysis and the option to use automatic confluency email alerts, can be applied to inform the user when certain culture conditions are reached (for example, once the cell culture has reached the desired confluency). Hence, the CytoSMART Lux2 can be used in many different ways to facilitate cell culture work and research.
In the following appnote several examples of applications of the CytoSMART Lux2 will be shown.
- Culturing cells in hypoxic conditions
- Standardizing cell culturing conditions
- The effect of confluency on transfection efficiency