The Erasmus MC iPS core facility

 

Welcome.

The Erasmus MC iPS core facility has been founded to support fundamental and translational research by providing high quality iPS cell lines and embryonic stem (ES) cell lines to researchers within and outside the Erasmus MC. For Erasmus MC employees, the services of the facility are available at subsidized costs.

The potential for iPS cell-related technology in regenerative medicine is widely appreciated and iPS cell research represents one of the fastest growing research fields in the world. IPS cells can be genetically engineered and differentiated into virtually any tissue or cell type and can therefore be used to understand human disease without the requirement of the diseased tissue. In addition, differentiated iPS cells can be used for drug screening and toxicology studies. Moreover, in the long term, differentiated iPS cells will be used for transplantation purposes.

The iPS core facility has ample experience in the generation of iPS cells. From many patients and controls, iPS cell lines have been generated and delivered to researchers within and outside the Erasmus MC. These high quality iPS cell lines are generated by experienced researchers, applying cutting edge technology according to standard operating procedures. Besides the generation of iPS cells, the iPS core facility also provides hands-on iPS and ES cell culture training and generates and provides mouse embryonic fibroblasts (MEFs) and cell culture reagents.

Erasmus MC iPS Core Facility has a METC approval named as 'Creation of disease model systems to understand and correct genetic disease through gene or other therapy using iPS cell derived from somatic cells: IPSC protocol Rotterdam'

Erasmus MC researchers that are willing to generate new iPSC lines from control and patient material, can use this protocol. Please contact manager of iPS Core Facility, Mehrnaz Ghazvini ( m.ghazvini@erasmusmc.nl) for more information.



  • slide show

    Derivation services

    The Erasmus MC iPS core facility provides all services needed to generate iPS cell lines, starting with skin biopsies, cultured fibroblasts, erythroblasts or bone marrow. Please find an overview of the service modules and fees.

  • cell culture support

    Culture support

    In case of difficulties with irradiated MEFs, culturing iPS cells, providing medium, the experienced staff of the iPS core facility can offer you culture support. Please contact Mehrnaz Ghazvini for information.

  • Request

    Request procedure

    Contact the technical director, Mehrnaz Ghazvini, m.ghazvini@erasmusmc.nl. The feasibility of your request will be determined during a personal intake. After the intake, you will be asked to fill in an agreement.

Services

Derivation agreement, protocols.



Category Service modules   Other institutes
Derivation services Module 1:
Grow dermal fibroblast from skin biopsy
   
Module 2a:
Reprogramming fibroblasts using lentiviral method
  Contact M. Ghazvini
Module 2b:
Reprogramming erythroblasts using lentiviral method
  Contact M. Ghazvini
Module 2c:
Reprogramming bone marrow(erythroblasts) using the lentiviral method
  Contact M. Ghazvini
Module 3:
Growing existing iPS cells for expansion or collecting other material
   
Failure reprogramming (two attempts)    
Culture support service Irradiated MEFS, Wt and DR4 Mail to: ipscore@erasmusmc.nl
Training Workshop pluripotent stem cell culture    








Module 1:
Grow dermal fibroblasts from skin biopsy.

This service takes up to one month. Fresh biopsies (2-4 millimeters) will be accepted to establish primary fibroblast line. From each biopsy we freeze several vials between passage 1 to 3 depends of growth ability of the cells.
Please note that improper biopsies or shipping of samples might result in an inability to grow fibroblasts or unsuccessful reprogramming.

Request procedure

Contact the technical director, Mehrnaz Ghazvini, m.ghazvini@erasmusmc.nl.
The feasibility of your request will be determined during a personal intake. After the intake, you will be asked to fill in an agreement.

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Module 2a:
Reprogramming fibroblasts using lentiviral method.

This service takes three months to complete and includes:

Fibroblast expansion, mycoplasma testing, freezing vials and plating for transduction.
Transduction of 1 well (100,000 cells/well) with lentivirus OSKM-dTomato.
Seeding transduced fibroblasts on irradiated MEFs. The rest of the transduced cells will be frozen.
Feeding every day.

  • Picking 6-12 colonies about month later
  • Expansion of colonies. 6 of them will be fully characterized.
  • Freezing 6 vials per line between passage 6 to 9. In some cases, due to reduced grow ability of some lines, we have to grow them to higher passages.
Characterization include:
  • RNA extraction, cDNA synthesis and qRT-PCR for endogenous Oct4, Nanog, Sox2, Cmyc, Gdf3, Rex1, Fgf4, Esg1, hTERT, Klf4;
  • Embryoid body formation at day 8, RNA extraction, cDNA synthesis and qRT-PCR for mesoderm markers Flk1 and GATA2, endoderm marker AFP, and ectoderm marker Pax6;
  • Immunofluorescence for Oct4, Nanog, TRA-1-81, SSE4;
  • Immunofluorescence for GFAP (ectoderm), Vimentin (mesoderm) and AFP (endoderm)
Chromosome count
Mycoplasma test of all the clones
Freezing of 6 vials per clone

Request procedure

Contact the technical director, Mehrnaz Ghazvini, m.ghazvini@erasmusmc.nl.
The feasibility of your request will be determined during a personal intake. After the intake, you will be asked to fill in an agreement.

×

Module 2b:
Reprogramming erythroblasts using lentiviral method

This service takes three months to complete and includes:

  • Growth of erythroblasts, mycoplasma testing, freezing of vials and plate for transduction
  • Transduction of cells with lentiviral vector
  • Picking 10-12 colonies (if possible)
  • Expansion and characterization of 6 colonies
  • Characterizations include:
    • Checking of pluripotency markers and markers of embryonal body differentiation at RNA level (RT-qPCR) and protein level (immunofluorescence)
    • Mycoplasma testing
    • Karyotyping
  • Freezing 6 vials per clone

Request procedure

Contact the technical director, Mehrnaz Ghazvini, m.ghazvini@erasmusmc.nl.
The feasibility of your request will be determined during a personal intake. After the intake, you will be asked to fill in an agreement.

×

Module 2c:
Reprogramming bone marrow (erythroblasts) using lentiviral method.

This service takes three months to complete and includes:

  • Growth of erythroblasts, mycoplasma testing, freezing of vials and plate for transduction
  • Transduction of cells with lentiviral vector
  • Picking 10-12 colonies (if possible)
  • Expansion and characterization of 6 colonies
  • Characterizations include:
    • Checking of pluripotency markers and markers of embryonal body differentiation at RNA level (RT-qPCR) and protein level (immunofluorescence)
    • Mycoplasma testing
    • Karyotyping
  • Freezing 6 vials per clone

Request procedure

Contact the technical director, Mehrnaz Ghazvini, m.ghazvini@erasmusmc.nl.
The feasibility of your request will be determined during a personal intake. After the intake, you will be asked to fill an agreement.

×

Module 3:
Growing existing iPS cells for expansion or collecting other material.

Growing existing iPS lines, expansion, freezing and delivering cells in culture

Request procedure

Contact the technical director, Mehrnaz Ghazvini, m.ghazvini@erasmusmc.nl.
The feasibility of your request will be determined during a personal intake. After the intake, you will be asked to fill an agreement.

×

Failure reprogramming (2 attempts).

In very few cases, reprogramming might fail. In such cases we will repeat the reprogramming one more time. In case of repeated failure, the two failed attempts will only be charged partially.

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Workshop

During this course precipitants will learn how to thaw, split and freeze pluripotent stem cells.
Also participants receive information about growing pluripotent stem cells on feeders versus feeder free culture system.

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iPS lines

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