StemScale™ PSC 悬浮培养基

The StemScale StemScale PSC Suspension Medium passaging protocol recommends passaging every 4-5 days, depending on spheroid size. We recommend passaging suspension cultures when the average spheroid diameter is 400 µm, which occurs approximately after 4-5 days of growth. Passaging cultures on day 3 is also an option but may result in a lower cell yield. Day 3 passaging can be performed to avoid passaging on a weekend.

Passaging protocols for other suspension culture media may offer less flexibility, depending on whether they utilize a fed-batch or overlay strategy. Generally, these cultures require a strict passaging schedule with little room for flexibility on the weekend.

The StemScale PSC Suspension Medium passaging protocol does not require the use of microcarriers or cell strainers, while other PSC suspension culture medium protocols do

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The StemScale PSC Suspension Medium feeding method involves replacing 50% of the spent medium with fresh medium. This feeding method prevents spheroids from being cultured in a medium that is accumulating significant quantities of waste by-products.

Feeding methods that use a fed-batch or overlay strategy do not remove the spent medium from suspension cultures. This method reduces hands-on time when feeding cultures; however, the accumulation of waste by-products is likely to negatively impact the health of these cells.

The StemScale PSC Suspension Medium feeding strategy also ensures a consistent volume through the growth period of the PSC spheroids. This is in contrast to batch-feeding methods where the volume increases incrementally. Since culture volume is one of the determinants of shear force and spheroid size, the 50% feeding strategy maintains this variable constant throughout the growth period.

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While other suspension culture media may require daily feeding of suspension cultures, the StemScale feeding schedule allows for every-other-day feeding. With StemScale PSC Suspension Medium, cultures can be fed daily to maximize cell health. However, users have the option to feed every other day without sacrificing performance while also providing more flexibility in the suspension culture schedule.

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The StemScale PSC Suspension Medium kit consists of two components: a basal medium and a supplement. Following reconstitution, the complete medium is used for all steps, from initiating cultures to feeding cultures. Other suspension culture medium kits sometimes consist of multiple components, which may be utilized at different points in the suspension culture process.

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Without DNase, the presence of small quantities of lysed DNA due to dissociation by TrypLE Select Enzyme will cause some spheroids to aggregate into a single clump.

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