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We offer protocols for the implementation of Corning Dissolvable Microcarriers (DMC) as a means to save time as you get started. We also providetechnical support resources for additional guidance.
We typically advise a process be developed in stages: Start by performing studies with your cells and media on microcarriers in ultra-low attachment multi-well plates to assess attachment to the microcarrier. Studies in spinner flask format can provide an early read on attachment and media conditions for agitated culture. For instance, a spinner flask may indicate if serum concentration needs to be lowered for better cell attachment in the dynamic attachment phase and later increased once attachment is sufficient. Spinner flasks also allow for assessment of the effect of nutrient consumption and metabolite production on cell growth and provide insight into subsequent feeding and/or media exchange requirements. In bench top bioreactors, the initial focus should be on the cell attachment phase and optimizing conditions such as agitation and plant densities in microcarrier culture. We recommended that you operate your bioreactor at an agitation rate that simplysuspends the microcarriers, in order to minimize shear and increase agitation. Consider the use of Poloxamer 188 to help mitigate stresses associated with bioreactor culture. From there, monitor cell growth as compared to static conditions and adjust conditions to maximize growth. Many cell types, including hMSCs, can be propagated on DMC in a bead-to-bead fashion allowing for cell expansion by successive additions of DMC and media, which allows cells to naturally migrate to the new surface areas. This method streamlines the cell expansion phase with less manipulation of the culture. Once you have reached your biomass production, the cell harvest protocol can be followed to gently dissolve the DMC for simple and efficient harvest of cells for downstream use.
Prior to investing too much time in bioreactor process development, we recommend working backwards from your desired production volume. . More specifically, when considering your target manufacturing scale, you can estimate an annual production rate. For example, if you find that you must run your process at 1 g of DMC per liter of media vs. 3g/L in order to meet your needs, this will greatly inform how you start the development of your process and possibly reduce false starts. While all bioreactor cell culture requires an investment in development time,timelines can be shortenedwith Corning’s technical support,.