Placenta – amnion
Evercyte ́s P-MSC/TERT308 cell line has been developed from amniotic membrane-derived mesenchymal stromal cells (placental-derived amnion), can be grown without limitations, shows typical markers and functions of MSCs and therefore represents a valuable model for studying processes such as differentiation, inflammation, tissue homeostasis and repair. In recent years evidence has accumulated that the potency of mesenchymal stem cells at least in part is mediated by secreted extracellular vesicles (EVs). P-MSC/TERT308 cell line has been established under conditions that will allow production of EVs for clinical applications.
Morphology and growth characteristics
P-MSC/TERT308 cells can be grown for a minimum of 50 population doublings with a stable growth rate and without showing signs of growth retardation. The cells are characterized by the typical spindle-shaped mesenchymal morphology.
Expression of MSC marker proteins CD73, CD90, CD105
P-MSC/TERT308 cells express typical marker proteins of mesenchymal stem cells such as CD73, CD90 and CD105, whereas the hematopoietic stem cell marker CD34 is not expressed (green peaks). Cells stained with isotype control antibodies are used as negative control (red peaks).
In vitro propagation
MSC NutriStem® XF medium + G418
MSC NutriStem® XF Basal Medium (Biological Industries / SATORIUS, Cat# 05-200-1)
MSC NutriStem® XF Supplement Mix (Biological Industries / SATORIUS, Cat# 05-201-1)
Additional material & reagents
Phosphate buffered saline (PBS) (Sigma, Cat# D8537)
CTS TrypLE Select Enzyme (Gibco, Cat# A1285901)
Passaging of cells
The new culture flasks have to be pre-coated with NutriCoat™ Attachment Solution (Biological Industries/SATORIUS, Cat# 05-760-1-15) following the instructions of the manufacturer. Briefly, dilute the substrate 1:500 in Ringer Solution and transfer the diluted substrate to the culture flasks (72 µl/cm²), incubate at least 1 hours at 37°C.
CryoStor® cell cryopreservation medium CS10 (Sigma Aldrich, Cat# C2874)
Additional material & reagents
Freezing of cells
Detach the cells after having reached about 70-80 % confluence from the culture vessel by using CTS TrypLE Select Enzyme solution (Protocol passaging of P-MSC/TERT308).
Thawing of cells
Original Evercyte cells are to be thawed in a T25 roux flask.
The business concept of Evercyte is to out-license telomerized cells to our customers. The license conditions depend on whether the contract partner is a for profit or a nonprofit organization and the intended use of the cells.
On time payment for unlimited use: EUR 1600
Pharmaceutical – chemical- cosmetic industries
Contract research organizations (CRO)
Initial license fee for 3 months: EUR 2500Annual license fee R&D: royalty based
production of extracellular vesicles using hollow fiber bioreactor
“I have had the pleasure of working with Evercyte for the last few years. We continually rely on Evercyte because of the high-quality data that they produce, their diligent responsiveness, and their excellent customer service.”
Josh Garlich, Senior Research Scientist, Apellis Pharmaceuticals, Inc.
“Cytonus has been working with Evercyte from many years as they are a trusted partner and have always delivered the highest quality cell lines to advance our platform. We routinely draw on their expertise to meet cellular engineering challenges and they have not disappointed.”
Remo Moomiaie-Qajar, Cytonus Therapeutics, Inc.