January 1, 1970 (Vol. , No. )

Steve Fiene Cellular Dynamics International
Giorgia Salvagiotto, Ph.D. Cellular Dynamics International

Familiarity with and careful attention to culture conditions is imperative to ensure cellular properties and functions are maintained over time.

Human iPSC-derived cells are not difficult to culture; they simply require familiarity and careful attention to the culture conditions to ensure that cellular properties and functions are maintained over time. Here are eight tips to enable successful applications of iPSC-derived cells:

1. Pick your human source based upon your research needs. Human induced pluripotent stem cell (iPSC)-derived cells enable researchers to select in vitro cellular systems generated from panels of healthy or diseased individuals, representing the genetic diversity within a given population.

2. Understand that the culturing environment can impact cellular functionality. Culture conditions are critical for maintaining cell health or inducing cellular stress. Some iPSC-derived cells require time to adjust to their environment after plating.

3. Select your culture medium based on your intended assay. Different cell types can have different metabolic and functional needs. Customized energy sources and medium supplements can modulate a cell’s function in the desired endpoint assay. A medium formulation different from the culturing medium may be required for optimal performance of the desired endpoint assay.

4. Determine the optimal timing for an assay based on the cellular function to be tested. Some cellular functions of iPSC-derived cell types require a degree of inter-cell communication, which is established over time. Examples of such time-sensitive cellular activities are neurite outgrowth, cardiomyocyte sarcomeric and gap-junction organization, and endothelial barrier function.

5. Select the optimal extracellular matrix (ECM), as it impacts cell adherence and functionality. Although in some cases several ECMs can allow cell adherence, optimal ECMs vary by cell type and can impact inter- and intra-cellular organization, cell functionality, and long-term culture.

6. Do not attempt to economize cells at the plating step, as it will impinge on the final assay results. Cell plating density can drastically affect gene and protein expression as well as cell functionality. Special attention to seeding procedures is particularly required for post-mitotic iPSC-derived cell types to achieve the appropriate cell densities that maintain cell functionality over time.

7. Pay special attention to the quality and quantity of cell recovery at the dissociation step to preserve the functionality of the iPSC-derived cells for subsequent assays. Dissociation enzymes, treatment time, and cell trituration procedures can affect the cell performance post-dissociation and the success of the endpoint assay.

8. Optimize your assays when using iPSC-derived cells. Just as any assay with a common cell line will need optimization, so too will assays with iPSC-derived cells. Simple changes to the culturing conditions can often overcome obstacles.

Effect of Extracellular Matrix Protein on iPSC-derived Neurons’ Morphology. iPSC-derived neurons (iCell® Neurons, Cellular Dynamics International) were cultured for 2 days on different extracellular matrix proteins: (a) Poly-D-lysine (pre-coated) overlaid with Matrigel (freshly coated); (b) Poly-L-ornithine (freshly coated) overlaid with laminin (freshly coated); (c) Matrigel (freshly coated); (d) Poly-D-Lysine (pre-coated). Images were taken at 100X total magnification.

Steve Fiene ([email protected]) and Giorgia Salvagiotto, Ph.D., ([email protected]) are application support scientists at Cellular Dynamics International.

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