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Services

Service Cost *
Isolation of fibroblasts from skin biopsy $500.00
Isolation of PBMCs from peripheral blood $100.00
Basic reprogramming/patient (3 clones/2 vials each) $7,500.00
iPSC characterization (ICC, gene expression) $1,450.00/line
Karyotyping $750.00/line
DNA fingerprinting $450.00/line
Mycoplasma test (required) $175.00
Expansion and banking on site $1,500.00/line

* Outside investigators and commercial entities please contact the Core Manager Marianne mfjames@bu.edu

How to request a line:

Recipients of iPSC lines outside of Boston University need to execute a Material Transfer Agreement (MTA) prior to delivery.

To request a line follow these steps:

  1. 1. Send an email to the iPSC Core Manager Marianne James (mfjames@bu.edu) listing the iPSC lines you are requesting.
  2. 2. Fill out the Material Transfer Agreement (MTA) Request Form, completing sections 2, 3 and 4 (and only sections 2, 3, and 4; leave all the rest unchanged please). Click the “Save” button at the end of the of the document, and email the form to Marianne James (mfjames@bu.edu). Do not click the “Submit” button.
  3. 3. The Boston University Office of Technology Development (OTD)/signing officials will then contact your OTD/signing official with an MTA document that will need to be signed by both your institution as well as the Boston University signing official.

iPSC derivation from PBMCs using STEMCCA lentivirus or Sendai virus

For this service we require 8 ml of freshly collected blood in CPT tubes (BD Biosciences – Cat# 362760) or other heparinized or EDTA tubes. Since the blood sample should be processed in less than one hour from the collection, please contact the Core manager to coordinate the delivery. The investigator can also submit frozen PBMCs.

This service takes 2-3 months and includes:

  1. Isolation and cryopreservation of PBMCs from peripheral blood
  2. Expansion of erythroblasts and mycoplasma testing
  3. Transduction of erythroblasts with lentivirus or Sendai virus
  4. Plating cells onto mitomycin C-treated MEFs and feeding everyday
  5. iPSC colony picking
  6. Expansion of three iPSC clones
  7. Mycoplasma testing
  8. Cryopreservation of iPSC lines (2 vials/clone)
  9. Shipping of frozen vials to investigator

Isolation of fibroblast from skin biopsy

  1. Isolation of fibroblasts from freshly collected skin biopsy
  2. Expansion of fibroblasts and mycoplasma testing
  3. Cryopreservation of early passage fibroblasts for storage and RNA/DNA isolation

iPSC derivation from fibroblasts using STEMCCA lentivirus or Sendai virus

Instead of a skin biopsy, investigators may choose to supply fibroblasts in frozen vials or bring them to the core as live cultures. In such cases, the investigator is required to test the fibroblasts for mycoplasma before submission. Only samples that are negative for mycoplasma will be accepted. The fibroblasts should be proliferating and at a low passage (preferably less than 8). Reprogramming of senescent cells is inefficient.

This service takes 2-3 months and includes:

  1. Expansion and cryopreservation of fibroblasts
  2. Mycoplasma testing
  3. Transduction of fibroblasts with lentivirus or Sendai virus
  4. Plating cells onto mitomycin C-treated MEFs and feeding every 24 hours for 30 days
  5. iPSC colony picking
  6. Expansion of three iPSC clones
  7. Mycoplasma testing
  8. Cryopreservation of iPSC lines (2 vials/clone)
  9. Shipping of frozen vials to investigator

Characterization of iPSC lines

Immunostaining and gene expression analysis

This service takes 1-2 months and includes:

  1. Immunostaining for the pluripotency markers Oct4, TRA-1-81, SSEA1 and SSEA4
  2. RNA extraction, cDNA synthesis and Real Time qPCR analysis for the expression of endogenous Oct4, Nanog, Sox2, Rex1, hTERT and Dnmt3b

Karyotyping

This service takes about 1 month and includes:

  1. Expanding the iPSC line to a T25 flask
  2. Shipping the iPSC line to Cell Line Genetics for G-band karyotyping

DNA fingerprinting

This service takes about 1 month and includes:

  1. Adaptation of the iPSC line to feeder-free culture conditions and genomic DNA extraction
  2. Genomic DNA extraction from donor cells (fibroblasts or PBMCs)
  3. Shipping the genomic DNA samples to Cell Line Genetics for Short Tandem Repeat (STR) analysis

Expansion and banking on site

The Core can provide expansion and banking services, consisting of long-term storage (three years) of 10 vials of each iPSC line.