Cell lines FAQ

• Can I use your reporter gene cell lines in immunocompetent mice?

  Most of our cell lines contain reporter genes or selection genes that are immunogenic. Therefore, we recommend that most of our cell lines be used in immunocompromised mice to get the most consistent results. An exception is our monoclonal mNIS-expressing mouse cell lines, which can be used in syngeneic hosts. For a more detailed discussion of using reporter gene-expressing cell lines in immunogenic mice, read our Science Talk blog “Betting against immunology: Choosing the right reporter gene for your oncology models.”

• Have your cell lines been tested for metastases in vivo?

  Our cell lines undergo extensive in vitro quality control testing. Select cell lines are tested in vivo, typically by subcutaneous injection. Additionally, we have tested some select lines for growth and metastases in either immunocompromised mice or syngeneic hosts. In vivo tests of product lines are shown in the “Reporter Validation” tab of the product page.

• How many times can I passage your cells?

  A lot. Our cells are made by lentiviral vector transduction, which means that the transgenes are stably integrated into the host cell genome and are very stable over many passages. However, keep in mind that it’s always a good idea to make a low passage stock of cells that you can go back to should any issues arise. Also, most of our cell lines are polyclonal populations, so we recommend passaging them in the presence of selecting antibiotics in order to ensure continued high reporter gene expression. The antibiotics and concentrations to use are indicated in the “Growth Conditions” tab of the relevant cell line product pages.

• Have your cell lines been tested for replication competent lentivirus (RCL)?

  We do not test our cell lines for RCL. Institutional requirements for RCL testing vary. Check with your institutional biosafety office for more information. To learn more about RCL testing offered by Imanis click here.

• Have your cells been tested for mouse pathogens?

  Our cell lines are not tested for contaminating murine pathogens. The testing requirements (which pathogens to test for) often vary between institutes. By not performing such testing, we keep our cell lines priced lower. Each of our cell lines is tested for mycoplasma, which is commonly included as part of any PCR testing.

   

• What growth conditions are needed for culturing your cells?

  All of our cells should be grown at 37^oC/5% CO₂. The type of media and passaging requirements vary between our different cell lines. Specific growth conditions, including media compositions, are shown in the “Growth Conditions” tab of the relevant product pages.

• What is replication competent lentivirus (RCL)?

  Replication competent lentiviruses (RCL) are virus particles capable of infecting cells and replicating to produce additional infectious particles.

  The existence of RCL in stable cell lines generated by lentiviral vector transduction is a safety concern. As such lentiviral vectors have been engineered to significantly reduce the likelihood of RCL production. These engineered lentiviral vectors have an excellent safety record, as there are no known reports of actual RCL production.

  Although RCL remains a theoretical concern, most institutes have specific requirements related to testing transduced cells for RCL. A p24 ELISA assay is the most commonly used test to demonstrate the absence of RCL in a sample of transduced cells. Imanis offers p24 ELISA RCL testing for lentivirus; visit our Sample Analysis page for more information.

• What is the source of the transgenes?

  Our transgenes come from a variety of sources. Check out the table below for detailed information about the source of our transgenes.

  Transgene	Accession #	Gene Source	Description
  Luc2	AY738222	Photinus pyralis	Codon optimized firefly luciferase with brighter signal1.
  hNIS	U66088	Homo sapiens	Human sodium iodide symporter; mediates iodide uptake2.
  hNISplus	Mutant of U66088	Homo sapiens	Genetically modified hNIS with enhanced radiotracer uptake.
  mNIS	AF235001	Mus musculus	Murine (mouse) sodium iodide symporter; mediates iodide uptake3.
  rNIS	U60282.1	Rattus norvegicus	Rat sodium iodide symporter; mediates iodide uptake4.
  pigNIS	NM214410	Sus scrofa	Pig sodium iodide symporter; mediates iodide uptake5.
  RhNIS	N/A*	Rhesus macaque	Rhesus sodium iodide symporter; mediates iodide uptake.
  dNIS	XM_541946	Canis lupus	Dog sodium iodide symporter; mediates iodide uptake6.
  IRES	M81861 (nt 260 to 848)	EMCV	Internal ribosome entry site; RNA element that mediates internal translation initiation7.
  eGFP	AAB02572.1	Variant of GFP from Aequorea victoria	Enhanced GFP; mutant of GFP with a 100-fold increase in fluorescent signal8.
  iRFP	JN247409	Rhodopseudomonas palustris	Near-infrared fluorescent protein; red-shifted fluorescent protein with an excitation/emission of 690/713 nm9.
  dsRed	AB212907	Anopheles gambiae	Red fluorescent protein variant with an excitation/emission of 558/583 nm10.
  DRD2	NM_016574.3	Homo Sapiens	Dopamine receptor D2; endogenously expressed in the brain11.
  hNET	NM_001172501.1	Homo Sapiens	Human norepinephrine transporter; endogenously expressed by noradrenergic neurons12.
  SSTR2	AY236542.1	Homo Sapiens	Somatostatin receptor 2; endogenously expressed in cerebrum and kidneys13.
  HSV-TK	JQ352282.1	HSV	Human herpesvirus 1 thymidine kinase (UL23)14.
  hTYR	M27160.1	Homo sapiens	Tyrosinase; converts tyrosine to brown-pigmented melanin15.

  *Sequence derived from a cDNA library. Email support@imanislife.com for more information.
  ¹ Promega
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