Frequently asked questions (FAQs)
DIFFERENTIATED CELLS FROM CELLARTIS
DOES CELLARTIS PROVIDE CARDIOMYOCYTES?
Cellartis provides cardiomyocytes derived from hES cells. The cardiomyocytes are available in two formats, either as beating clusters, hES-CMC™002, or as beating monolayers, hES-CMC™2D. The cardiomyocytes express many cardiac markers and ion channels such as hERG, L-type calcium channels, voltage-gated potassium and sodium channels, and funny channels. The cardiomyocytes respond to known ion-channel blockers like E-4031, astemizole, verapamil, and lidocain and also respond to pharmacological stimuli such as α-Adrenergic and β-Adrenergic stimulations as well as muscarinic stimulation. The cardiomyocytes can be used for electrophysiological recordings and evaluations of drug-induced changes of the QT interval. The cells are non-proliferating and are supplied fresh within 6-8 weeks after order. Please see the product sheets for information or contact Cellartis.
DOES CELLARTIS PROVIDE HEPATOCYTES?
Cellartis provides hepatocyte-like cells derived from hES cells in 24w and 96w plate formats.
The hES-HEP™002 cells have hepatocyte morphology and express a number of hepatocyte properties like hepatic transcription factors, specific proteins, CYPs, transporters and specific enzyme activities. The hepatocyte-like cells stores glycogen and show CYP1A activity. The cells are supplied fresh within 6-8 weeks after order. Please see the product sheets for information or contact Cellartis.
UNDIFFERENTIATED hES CELLS FROM CELLARTIS
DOES CELLARTIS HAVE hES CELL ELIGIBLE FOR USE IN MY COUNTRY?
The company has been deriving hES cell lines since early 2001 and has cell lines that are eligible for use in all countries that permit hES cell research. Two of the cell lines are listed on the original National Institutes of Health (NIH) Human Embryonic Stem Cell Registry from 2001 and are thus eligible for US federal funding. For all other markets, Cellartis will provide you with detailed information about the available hES cell lines that can be used in your country and what type of funding they are eligible for.
HOW MANY DIFFERENT hES CELL LINES DOES CELLARTIS HAVE AVAILABLE?
Cellartis is the largest single source of defined hES cell lines worldwide and has more than 30 hES cell lines and sub clones available today.
CAN I OBTAIN hES CELLS IN EARLY PASSAGES FROM CELLARTIS?
Cellartis produces and provides hES cells in early passages (passage 11 and up). Late passages are also available upon request.
CAN I OBTAIN CLONALLY DERIVED hES CELLS FROM CELLARTIS?
Cellartis has created clonally derived hES cells from selected parental cell lines. These are available upon request.
HOW CAN I CHOOSE BETWEEN THE DIFFERENT hES CELL LINES FROM CELLARTIS?
Cellartis is available for personal consultation and advice on the selection of specific hES cell lines and passages in order to make sure that you receive optimal cell lines for your specific applications. The large pool of defined hES cell lines from Cellartis ensures the possibility of finding optimal cell lines for different applications.
WHERE CAN I FIND MORE INFORMATION ABOUT INDIVIDUAL hES CELL LINES FROM CELLARTIS?
The best way to get updated and detailed information about the hES cell lines from Cellartis is through direct communication with Cellartis representatives. In addition, other sources of information include the listings on the NIH Human Embryonic Stem Cell Registry, the Wisconsin International Stem Cell Bank, the ISSCR web-site and through scientific publications from Cellartis.
DOES CELLARTIS PROVIDE ADDITIONAL PRODUCTS THAT FACILIATE THE IMPLEMENTATION AND WORK WITH HES CELLS IN MY LABORATORY?
In addition to characterized hES cell lines, Cellartis collaborates with Vitrolife AB to supply associated culture media and stem cell cutting tools.
CHARACTERIZATION AND PROVEN PLURIPOTENCY
HOW DO I KNOW THAT THE hES CELLS FROM CELLARTIS ARE UNDIFFERENTIATED AND PLURIPOTENT?
In order to guarantee that the hES cells delivered from Cellartis exhibit the fundamental properties of undifferentiated hES cells, a LOT procedure is employed for hES cell production. Basically this means that the hES cell lines are propagated and expanded until a substantial number of vials (>100) can be frozen at one time. The simultaneously frozen vials of a specific cell line are referred to as one LOT. All characterization analyses of each LOT are performed on hES cells sampled from the frozen LOT. This procedure assures the quality and pluripotency of the cells delivered from Cellartis. Yet another advantage with the LOT procedure is that researchers repeatedly can receive new hES cells from the same passage and freezing day.
HOW ARE THE hES CELLS CHARACTERIZED AT CELLARTIS?
Each hES cell line derived at Cellartis is extensively characterized. The characterization includes stem cell marker analysis (SSEA-1, -3, -4; TRA-1-60, -1-81 and Oct-4), and genetic characterization (karyotyping and FISH). Furthermore, telomerase and alkaline phosphatase activities are analyzed and the cells are assayed for in vivo and in vitro differentiation potential. The cell lines at Cellartis have been cultured in vitro for extended periods and endured repeated freezing and thawing cycles, while retaining pluripotency and stable karyotypes. For further information about the characteristics of the cell lines from Cellartis please see Heins et al. (Stem Cells, 2004).
ARE THE hES CELL LINES FROM CELLARTIS TESTED FOR HUMAN VIRUSES AND MYCOPLASMA?
All hES cell lines provided by Cellartis have been tested negative for HIV types 1 and 2, Hepatitis B and C, Cytomegalovirus, Herpes Simplex Virus type 1 and 2, Epstein-Barr Virus, and Human Papilloma Virus. In addition, the hES cell cultures are also regularly tested for the absence of mycoplasma.
ETHICAL CONSIDERATIONS
WHAT IS THE SOURCE OF EMBRYOS USED FOR THE hES CELLS DERIVATION PROCESS AT CELLARTIS?
All hES cell lines supplied by Cellartis are derived from supernumerary embryos created for the purpose of assisted reproduction and donated with written informed consent from the patients. The donations did not result in any financial gain for the patients and no information about the embryo donors are accessible. Cellartis warrants that all research and handling of hES cells is based on careful ethical considerations and the company is guided by and follows all existing laws and regulations in the major markets.
HOW ARE hES CELL LINES DERIVED AT CELLARTIS?
Establishment of hES cell lines at Cellartis is performed according to the procedures described in detail in Heins et al. (Stem Cells, 2004) (Patent Pending).
PROTOCOLS AND TECHNICAL SUPPORT
HOW DO I HANDLE (CULTURE, MAINTAIN, THAW AND FREEZE) hES CELLS FROM CELLARTIS?
When obtaining hES cell lines from Cellartis you will receive an extensive manual for the culture, freezing and thawing procedures. The manual will be mailed once a material transfer agreement (MTA) is completed and signed and the full payment has been made.
DOES CELLARTIS OFFER TECHNICAL SUPPORT?
The highly experienced team at Cellartis provides technical support for customers receiving hES cells from Cellartis. Typically, the basic technical support is accomplished via e-mail and telephone communications. However, other solutions are also available in order meet the specific requirements from the hES cell customers.
CAN CELLARTIS HELP ME WITH SETTING UP MY hES CELL CULTURES?
In order to guarantee the successful implementation of hES cell culture in the customers’ laboratories, Cellartis can provide extensive technical support on different levels.
SUPPLY OF hES CELLS
WHAT FORMS DO I HAVE TO COMPLETE TO OBTAIN hES CELLS FROM CELLARTIS?
Human ES cells and associated products are immediately available from Cellartis. In order to obtain hES cells from Cellartis, you must first obtain all relevant approvals to conduct your research project using hES cells. In order to deliver hES cells, Cellartis requires a signed Material Transfer Agreement (MTA) (or Simple Letter Agreement for NIH funded researchers) and that all relevant import permits have been obtained. Cellartis will assist with this process where possible. The MTA or Simple Letter Agreement must be signed by the Principal Investigator and the relevant authorities.
HOW DO I OBTAIN hES CELLS FROM CELLARTIS?
Obtaining hES cells from Cellartis is easy. The process is initiated by forwarding a completed MTA to Cellartis. Your payment will be required prior to shipment. Once the necessary documents and payments are completed and received, the hES cell lines are shipped immediately with an overnight door to door transport.
HOW ARE THE hES CELLS DELIVERED?
Cellartis ships cryopreserved hES cells by World Courier door to door. The cells are transported in a dry shipper with encapsulated liquid N2 keeping the cells in the vapor phase of N2. The hES cells from Cellartis are frozen by vitrification in closed straws (Heins et al., 2004) (Patent Pending). The closed straws protect the cells during storage and transport. The vitrification method developed by Cellartis ensures a high percentage of cell recovery upon thawing.
HOW DO I HANDLE THE hES CELLS UPON RECEIPT?
Each hES cell order from Cellartis is accompanied by a detailed step-by-step manual that describes all procedures for handling and culturing of the cells. Upon receipt the cells should be transferred from the transport vessel to a liquid N2 tank for storage until you want to thaw the cells for subsequent culture.
WHAT RESEARCH CAN BE CONDUCTED USING hES CELL LINES FROM CELLARTIS?
Cellartis requires that you comply with all Governmental and Institutional Ethics Committee regulations relating to the use of hES cell in your country. The restrictions in using hES cell lines from Cellartis are detailed in the Material Transfer Agreement (MTA) under which the cells are provided. The cell lines must be used in compliance with the MTA. Note, you may not transfer hES cells from Cellartis to another researcher or commercial party without the written permission of Cellartis.
ARE THERE RESTRICTIONS ON PUBLISHING DISCOVERIES MADE WITH hES CELLS FROM CELLARTIS?
Researchers obtaining hES cells from Cellartis are free to publish discoveries made with these cell lines, although Cellartis asks researchers to acknowledge the source of their hES cells in scientific journals and papers.
HANDLING OF hES CELL LINES FROM CELLARTIS
HOW ARE THE hES CELLS FROM CELLARTIS PROPAGATED?
For routine manual expansion, the hES cells are cultured on mitomycin C treated MEF using VitroHES™ medium provided by Vitrolife AB (Göteborg, Sweden) supplemented with 10 ng/ml hrbFGF. The hES cells are passaged to plates with fresh medium and mEF by mechanical dissection using the “Stem Cell Tool” (Vitrolife AB). Established hES cell lines are routinely passaged every 4-5 days. The hES cell colonies are mechanically cut into pieces, 200 x 200 µm, and removed from the culture dish and transferred to a new culture dish with fresh medium and mEF. In the laboratories of Cellartis, hES cells are also propagated using various enzymatic methods.
HOW ARE THE hES CELLS FROM CELLARTIS FROZEN?
The hES cells from Cellartis are frozen by vitrification in closed straws (Heins et al 2004) (Patent Pending). The closed straws protect the cells during storage and transport. The efficient vitrification method at Cellartis ensures a very high percentage of cell recovery upon thawing. Experiments in the laboratories of Cellartis have demonstrated that viable hES cells are recovered from every straw prepared using this vitrification procedure.
MEDIA AND TOOLS
DOES CELLARTIS SUPPLY OTHER hES CELL ASSOCIATED PRODUCTS?
The successful propagation of undifferentiated hES cells is critically dependent on high quality consumables, culture media, tools and feeder cells. In order to meet these requirements Cellartis, in collaboration with Vitrolife AB, has developed two serum free, quality controlled, ready-to-use media for the propagation and handling of undifferentiated hES cells; VitroHES™, a culture medium for undifferentiated hES cells, and VitroPBS™, a medium for the freezing and thawing of hES cells. Cellartis strongly recommends customers to initiate their hES cell cultures using VitroHES™. Vitrolife AB also provide stem cell cutting tools which have been designed for the cutting, manipulation and transfer of hES cell colonies. The Stem Cell Tools are non-toxic and offer uniformity, efficacy and safety. The tools are extremely durable and can be utilized during the whole cutting session without the need for replacement.
CAN hES CELLS FROM CELLARTIS BE CULTURED ON OTHER TYPES OF FEEDER CELLS OR IN FEEDER FREE CONDITIONS?
Cellartis have long experience with setting up and optimizing different culture conditions for hES cells. In the laboratories of Cellartis, hES cells are successfully cultured on human feeder cells and in feeder free culture systems. However, Cellartis strongly recommends customers to initiate their hES cell cultures using recommended mEF provided by ATTC.