Cell culture on microcarriers is an increasingly important technique for the production of large cell quantities and the scale-up of Cell Therapy applications. Microcarriers—small, spherical beads used to grow adherent cells in a suspension culture—offer several advantages for large-scale cell culture: Because they are suspended, microcarriers allow for a maximized surface area-to-volume ratio. Single-vessel conditions make microcarriers amenable to scale-up applications. Microcarriers also offer the ability to monitor and control physiological conditions when used in bioreactors.
As cell therapies become more common in clinics, there is need for more streamlined scale-up, process robustness, cost efficiency, and regulatory compliance. To meet this need, Corning® has developed a new generation of microcarriers which can be dissolved with a solution of EDTA and pectinase. This new dissolvable microcarrier technology allows cells to be recovered without the need for microcarrier separation. This gentle cell harvest is more ideal than traditional microcarriers for Cell Therapy applications.
This Ask the Expert session will aim to answer all of your questions about dissolvable microcarriers as a next generation technology for stem Cell Therapy.
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The advantages of moving into microcarrier based culture can be realized at relatively small scale. Operation of a bioreactor with a 2.5L working volume using 2 g/L of Corning Dissolvable Microcarriers equates to approximately 25000 cm² of available surface area for cell culture while occupying a small footprint of bench space. The equivalent to this … Continued
In my research on microcarrier use, I have read about optimization taking a long time. Do you have any advice on streamlining this process and how does is your product in terms of required optimization?
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 … Continued
Have you looked at whether the disposable material or the products used to initiate the dissolve effect the expression of surface markers morphology, or pluripotentcy of MSCs
Appropriate surface marker morphology and maintenance of pluripotentcy is certain to be of interest for a user of Corning Dissolvable Microcarriers (DMC) when producing stem cells. Understanding this, we evaluated expected surface markers, post-harvest pluripotentcy, and karyotype of stem cells across a multitude of different conditions: Our studies showed that this is true for both … Continued
Can you explain about the formation of cell aggregates with microcarriers, have you seen this and do you have any advice on how to limit this?
The formation of multiple microcarrier and cell aggregates is a common phenomenon seen for many, if not all, microcarriers under certain conditions. This is driven by cell adhesion molecules (CAMs) that are typically expressed as a consequence of growth and allow cells to adhere to extracellular matrix or other cells. As such, the extent of … Continued
Which types of bioreactors have you used the dissolvable microcarriers in? Specifically have you used these in wave type bags?
We are aware of the use of Corning Dissolvable Microcarriers (DMC) in various bioreactor platforms. Both glass and single use spinner flasks have been used extensively for small scale development work. We have seen DMC used in Eppendorf BioBLU (3c, 14c, and 50c), Millipore Sigma Mobius®, PBS Biotech® Vertical-Wheel™, and Sartorius BIOSTAT B systems. In … Continued
There are no special media supplements required for using Corning Dissolvable Microcarriers (DMC): The DMC have been used in an array of serum-containing, serum-free, and chemically-defined media. However, there are some considerations to keep in mind. First, cell culture media with a high metal ion content may require harvest condition optimization, as the dissolution harvest … Continued
Corning Dissolvable Microcarriers (DMC) are sufficiently durable to support multi-day culture, as we determined after fifteen days of cultivation to reach our biomass target. There was no indication that structure or function was negatively impacted at this time point. In terms of physical durability, it is worth nothing that, if DMCs were to become fractured, … Continued
The harvest procedure is available as a detailed resource document that your local Corning sales representative can provide. Essentially, once cells are ready for harvest, a wash step is performed with DPBS to rinse away culture media. This is often performed by settle-aspirate operation. Since Corning Dissolvable Microcarriers (DMC) are made of polygalacturonic acid (PGA) polymer … Continued
Cell attachment to Corning Dissolvable Microcarriers is quite efficient: We typically see greater than 90% attachment as determined by the count of free cells in the supernatant. There is a significant range of time required to attach for various processes, however. The characteristics of cell type and/or media used can impact how readily attachment occurs. … Continued
We have demonstrated successful growth with many cell types that are used in Cell Therapy applications, as well as standard bioprocess production cell lines. The advantages offered by Corning Dissolvable Microcarriers (DMC) are not limited to a particular cell type but are most valuable for specific applications. Due to the fast time to harvest and … Continued