Regenerative Medicine

From Cell Culture to Cell Manufacturing

Cell culture/manufacturing to produce a therapeutic product is very different than cell culture as routinely performed in an academic or basic research setting. Manufacturing in the healthcare and pharmaceutical industries requires a greater level of quality control and operational efficiency and is subject to significantly more regulation. Predictable production of a quality product that meets the required standards while minimizing cost is the ideal.

The application of image analysis to inform decision-making helps remove one of the greatest barriers towards predictable manufacturing – the reliance on human operators, which inherently introduces variability, even with an experienced staff. Qualitative methods, e.g., “shaking the vessel three times after seeding” can be put to the test using quantitative image analysis techniques and thus formalized.

Application of engineered 3D stem cell-derived systems for patient implantation is already happening, e.g., the culture of skin grafts. Clinical testing of more complicated cell systems is underway. The eventual goal is to be able to culture fully functional complex organs such as the heart. Note that 3D engineered tissues, including various types of organoids, are also applied towards drug discovery testing.

Minimizing Perturbation to Sensitive Cell Cultures

A variety of cell types are cultured in vitro, including primary cells recently obtained from tissue, immortalized cell lines, and stem cells. Cultures of immortalized cell lines, as often used for basic biological research, can be quite capable of withstanding significant insult, such as improper seeding density, being left outside of the incubator for extended periods, etc. Stem cells tend to be more sensitive to perturbation. Stringent attention to culture conditions is thus important when working with stem cells, especially when cultured for use as part of a therapeutic.

A top factor to consider is the incubation conditions, and consistency thereof. Nikon's BioStudio-T and BioStudio-mini are instead manufactured to be compact enough to fit in existing incubators and biosafety cabinets.

Vibration is another concern. Among other factors, vibration can cause cells stress and influence their seeding pattern following passage. To minimize vibration, the BioStudio-T uses an objective-scanning strategy for tiled acquisition, rather than moving the sample.

Phototoxicity is also a concern. For this reason, a low illumination intensity technique such as phase contrast is usually favored. Fluorescence labeling can be an invasive procedure and fluorescence imaging can be phototoxic. Nikon’s BioStudio series instruments take advantage of phase contrast imaging using a red LED for illumination. Red light carries less energy and is less phototoxic than a similar quantum of blue light.

Cell x Image Lab – A Resource for Cell Culture Success

Supporting the regenerative medicine field is a strategic goal of Nikon Healthcare. To this end, we have launched the Cell x Image Lab website, which details how microscopy imaging and analysis can be used to give cell culture processes and procedures a quantitative edge – enhancing the success rate and reproducibility of cell manufacturing. Imaging and analysis are also useful for quality assurance and developing new processes and procedures.

The Cell x Image Lab website includes both an introductory section with basic information about the field as well as numerous case studies detailing specific imaging and analysis applications for cell culture and manufacturing.