Stem Cell Culture Advances – The Final Blog in the ISSCR Series
In June I attended the International Society for Stem Cell Research (ISSCR) in Vancouver and began a series of blogs related to the information presented at the meeting. In today’s blog I will be covering novel technologies for large scale stem cell manufacturing, laminins in stem cell culture and smart incubators. The daily Innovation Showcases were particularity interesting and focused on new, state of the art products or approaches that enable stem cell culture and research.
Large Scale Stem Cell Manufacturing
As more stem cell therapeutics progress through clinical testing, in vitro culture methods must be scalable to deliver the quantities of cells needed. Unfortunately, current approaches can be cumbersome to scale. In the Innovation Showcase, “Expansion and Harvest of Adult Stem Cells Supports Large Scale Manufacturing,” Julie Murrell, Ph.D., R&D Head, Stem Cell Biology Lab, EMD Millipore, demonstrated an expansion paradigm that uses a scalable, single use, stirred tank bioreactor [1] with microcarrier scaffold for human mesenchymal stromal/stem cell (hMSC) culture.
This approach enables direct monitoring for the specific characteristics of hMSCs at any point during the culture, thus assuring product quality and consistency. The bioreactor provides ease of use in handling and reduced medium volume requirements [2]. In this case study, Dr. Murrell described full expansion to over 400 million cells, along with harvesting and characterization for hMSCs. Her team evaluated multiple harvest technologies and found that they provide greater than 90% recovery of high viability cells. Finally, the cells were evaluated using flow cytometry to assess a variety of markers that confirm identity and purity [3]. Differentiation potential was confirmed using traditional methods. Due to the limited ability to apply traditional biosafety steps in the downstream process for cell therapies, use of high quality reagents at every step is essential [4].
In this work, they verified that cells expanded in the single use stirred tank bioreactor and then harvested were identical in phenotypic profile to flat culture and maintained the desired cell characteristics of hMSCs. Which confirmed the consistency, quality, reproducibility and cost effectiveness of large scale in vitro systems for stem cell expansion.
4 http://www.emdmillipore.com/INTL/en/support/regulatory-support/emprove/9qmb.qB.T_gAAAFDRclUTz7y,nav
The Laminin Protein Family – key to primary cell culture
Dr Kristian Tryggvason gave a presentation on the novel BioLamina platform with cell-type specific human recombinant laminins that have unique physiological relevance that is of critical importance for stem cell culture. Researchers in the areas of regenerative medicine, basic biomedical research, and drug discovery all share a common need for reproducible procedures to culture human pluripotent stem cells and/or differentiated cells. By successfully recreating specific in-vivo cell niches in the dish, the talk described how recombinant laminins can 1) solve in essence all practical problems of expanding of pluripotent cells, 2) keep adult and differentiated cells in their differentiated state, 3) differentiating stem cells on biorelevant matrices.
The talk described how the biologically relevant laminin-521 matrix enables large-scale generation of hPSCs of clinical quality. Cells can be clonally and single-cell expanded without detrimental ROCK inhibitors and provides a high-quality system with genetically stable cells at high cell yield. BioLamina is the only company in the world that has succeeded in making full-length human recombinant laminins at a commercial scale and some of the new data presented at ISSCR were:
- Weekend-free Feeding – a protocol for completely weekend-free feeding.
- Transfer, expansion, clonal cultivation and EB differentiation of hPSCs on Laminin-521, Nature Protocols
- Articlesized, single-cell passaging without ROCKi and clonal derivation of human ECS in a completely xeno and feeder free environment, Nature Communications
A compelling case was made for the use of recombinant laminins to enable robust derivation, expansion, and differentiation of cells in physiological microenvironments suitable for regenerative medicine and drug discovery.
“Smart” Incubators for Improved Culture
This year Panasonic launched their new line of cell IQ incubators with touch screen technology. At ISSCR, I was able to go by their booth and get a tour of the new line of smart incubators. The incubators have been specifically designed with different models to meet specific research needs. They have also employed several innovative new technologies to improve incubator performance. There are too many technologies to describe in the space of this blog, but I have described a few of my favorite features below:
Panasonic has incorporated a dual detector infrared CO2 sensor that provides extremely accurate readings. This enhanced accuracy allows the incubator to recover faster without overshoot even with multiple door openings by the researchers. Panasonic states that CO2 recovery can be achieved in as little as two minutes after the door closes.
Panasonic has also taken an innovative approach to contamination with both proactive and reactive technologies. In an effort to be proactive, the incubators have a copper-enriched stainless steel alloy interior surface, which functions as a natural germicide called InCu-saFE. On the reactive side they employ a decontamination technology based on H2O2 vapor, which has been shown to be more effective even on difficult to detect contaminants like mycoplasma. In addition, the time to decontaminate is reduced to just 3 hours.
The incubator also offers Active Background contamination control with IncuSafe and UV decontamination technology. This feature provides Proactive contamination control, even when cells are being cultured inside the incubator, thus providing a 24X7 protection from contamination.
Also, since the new design of this incubator allows for a significant reduction in movable and removable parts, the incubator is extremely easy to maintain, leading up to 80% savings in maintenance time.
ISSCR Series
This is just the second in a series of blogs covering ISSCR 2014. It was an excellent meeting and I highly recommend attending if you get the opportunity. Find out more about the ISSCR’s 2015 Annual Meeting in Stockholm, June 24-27 by visiting – ISSCR’s 2015 Annual Meeting
ISSCR Series Blogs
“Innovations Abound at the 2014 Annual International Society for Stem Cell Research (ISSCR) Meeting”
“Progress is Being Made Toward Using Cells as a Therapy for Type 1 Diabetes”
“Cell Therapy for Parkinson’s Disease – Considerations for the Future”