CReM — Cores & Protocols
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iPSC Core Facility

Induced Pluripotent Stem Cells (iPSCs) generated by forced overexpression of defined transcription factors in somatic cells hold great promise for human disease research and personalized medicine. iPSCs show extensive self-renewal and have the ability to become any cell type in the body, providing an inexhaustible source of cells for in vitro disease modeling studies, screening of pharmacological compounds and regenerative therapies.
Tra1-81 staining of human iPSC colony
Tra1-81 staining of human iPSC colony
The Center for Regenerative Medicine (CReM) iPSC Core was created to expedite the use of iPSC technology by providing essential services and support to on-campus investigators and the broader scientific community. To achieve this goal the CReM iPSC Core will:
  • Generate and characterize iPSC lines from samples of adult tissues provided by the investigator. For more information visit services (for BU Faculty & Staff only).
  • Serve as a repository for sharing control and disease-specific hiPSC lines with internal and external investigators to conduct both basic and translational research. For more information visit our CReM bank.
  • Provide expertise, protocols and training to support work involving human iPSCs.

The CReM IPSC Catalog

The CReM iPSC Catalog is an interactive online catalog of induced pluripotent stem cells (iPSCs) that are available to the scientific research community upon request. This iPSC catalog was created by the Center for Regenerative Medicine (CReM) of Boston University and Boston Medical Center and allows users to view, inquire and order iPSCs from the CReM bank of control and disease-specific iPS cell lines including gene-edited and ‘knock-in’ reporter cell lines.

The CReM and CReM iPSC Core maintain a state-of-the-art bank of frozen iPSC lines that have been derived by CReM investigators and made readily available to all Boston University and external investigators. Downloadable iPSC characterization data are also available for most iPSC lines. The iPSC catalog is an automated ordering system that allows investigators to follow the approval status of their requests including Material Transfer Agreements (MTAs) and shipment scheduling.

Disease Specific iPSC biorepositories:

Interstitial Lung Disease (ILD)
Children’s Interstitial Lung Disease (chILD)
Creutzfeldt-Jakob Disease (CJD)
Sickle Cell Disease
Cystic Fibrosis
Alpha-1 Antitrypsin Deficiency
TTR Amyloid Disease (ATTR)
Framingham Heart Study (FHS)
Exceptional longevity (EL)-Specific iPSC Library

Our IPSC bank:

https://stemcellbank.bu.edu/Catalog/Item/Home

iPS Cell Colonies
iPS Cell Colonies

For inquiries concerning available iPS cells please contact us at mfjames@bu.edu

To request an iPSC line(s) please create an account by clicking Login tab (top right of main page) and follow the prompts and guidelines outlined on each page. Order status updates may be accessed through your “My Account” Orders History tab.

Note: Entries in the Search Catalog by Selection & Search Catalog by Keyword sections are combined together to form one search.

human iPSC training course
human iPSC training course

Human iPSC Training Course

Basic iPSC Hands-On Training Courses

The CReM iPSC Core is offering basic training in human induced pluripotent stem cell (hiPSC) culture for research investigators working with or planning to work with hiPS cells. The hands-on training will take place over 2 weeks (approximately 1 hour per day) and will provide investigators with the techniques necessary to maintain and propagate established iPS cells in their own lab. Training will focus on feeder-free (mTeSR1/Matrigel culture system) hiPS cell culture methods including colony passaging using common cell dissociation reagents and mechanical picking, identification of differentiated cells, cryopreservation and thawing. Detailed SOPs will be provided including methods for characterizing iPSCs for karytoype analysis and for pluripotency by immunofluorescent staining. Prior experience in general cell culture techniques is required.

This training is for the BU community only.

Please contact Marianne James for further information mfjames@bu.edu
Cost of Training Course: $1200/BU

Training Course in Human-Induced Pluripotent Stem Cells and Their Differentiation into Endoderm and Lung Progeny

This five-day course will focus on deriving, maintaining and characterizing pluripotent stem cells (hiPSCs) and their differentiation to endoderm and lung progeny. The course is designed for research scientists working with or planning to work with human iPSC culture who have prior experience in general cell culture techniques. It will include hands-on training, lectures, and demonstrations from leading experts and educators in the field of stem cell biology from the Center for Regenerative Medicine (CReM) at Boston University and Boston Medical Center. Small class size will enable researchers to learn the process of reprogramming from somatic cell preparations, including iPS cell identification, isolation and characterization, and current approaches in directed differentiation to endodermal and lung lineages. Additional topics will include lectures and training in novel gene editing techniques, including the design and use of CRISPR/Cas9 in iPSCs, and development of 3D culture systems.

This course is open for the entire scientific community. The course is temporarily suspended; no dates have been announced.

The in-person Training Course in Human-Induced Pluripotent Stem Cells and Their Differentiation into Endoderm and Lung Progeny was suspended during the pandemic and has moved to a remote learning format. You can access much of the content from this training course through our freely available documents, protocols and downloadable video tutorials at the following links:

Please contact Marianne James for further information mfjames@bu.edu

Services

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 is available for the BU Community Only.)

This service takes 2-3 months and includes:

  • Isolation and cryopreservation of PBMCs from peripheral blood
  • Expansion of erythroblasts and mycoplasma testing
    Transduction of erythroblasts with lentivirus or Sendai virus
  • Plating cells onto mitomycin C-treated MEFs and feeding everyday
  • iPSC colony picking
  • Expansion of three iPSC clones
  • Mycoplasma testing
  • Cryopreservation of iPSC lines (2 vials/clone)
  • Shipping of frozen vials to investigator
  • Isolation of fibroblast from skin biopsy
  • Isolation of fibroblasts from freshly collected skin biopsy
  • Expansion of fibroblasts and mycoplasma testing
  • 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 is available for the BU Community Only.)

This service takes 2-3 months and includes:

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

Characterization of iPSC lines

This service takes about 1 month and includes:

  • Immunostaining and gene expression analysis
  • Immunostaining for the pluripotency markers Oct4, TRA-1-81, SSEA1 and SSEA4
  • 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:

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

DNA fingerprinting

This service takes about 1 month and includes:

  • Adaptation of the iPSC line to feeder-free culture conditions and genomic DNA extraction
  • Genomic DNA extraction from donor cells (fibroblasts or PBMCs)
  • 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.

ServicesCost*
Isolation of fibroblast from skin biopsy$500.00
Isolation of PBMC's from peripherial blood$500.00
iPSC characterization (ICC, gene expression)$1,450.00/line
Karyotyping$750.00/line
DNA fingerprinting$450.00/line
Expansion and banking on site$1,500.00/line
Basic reprogramming/patient (3 clones/2 vials each)$7,500.00
Mycoplasma test (required)$175.00

iPSC Core Team

Rhiannon Darling
iPSC Core Technician
Trevor Koppy
iPSC Core Technician
Marianne James, PhD
iPSC Core Manager
CReM
email me

Biocomputational Core

The Biocomputational Core at the CReM is responsible for the design and analysis of all biocomputational studies currently ongoing in the different CReM labs. The Core uses different platforms and pipelines (WGS, RNA-seq, differential gene expression, single cell sequencing, DGE, etc) focusing on stem cell identity, differentiation into different lineages and the establishment of cellular state.

Biocomputational Core Team

Feiya Wang, MA
Bioinformatics Research Assistant
Feiya Wang
Pushpinder Bawa, PhD
Bioinformatics Core Manager
CReM
email me
Pushpinder Bawa

The Biocomputational Core at the CReM is responsible for the design and analysis of all biocomputational studies currently ongoing in the different CReM labs. The Core uses different platforms and pipelines (WGS, RNA-seq, differential gene expression, single cell sequencing, DGE, etc) focusing on stem cell identity, differentiation into different lineages and the establishment of cellular state.

We work together with our adjacent Boston University School of Medicine’s Single Cell Core, a facility containing state-of-the-art microfluidic and drop-seq based platforms for the capture, library preparation, and sequencing of single cells. More information available at: https://www.bumc.bu.edu/singlecell/

For internal use only: CReM Project Index (password required)

Protocols

As part of our mission of open-source biology, the Center for Regenerative Medicine (CReM) makes all of our protocols available to the entire scientific community.

Embryonic Stem Cell Culture Protocols

2i Media (Feeder Free, Serum Free)

iPS Cell Core SOPs

ESC/iPSC Directed Differentiation Protocols

Maintenance and freezing of iPSC-derived alveolospheres

iPS Cell Protocols

Vector Protocols

Other Protocols

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Welcome to CReM

My name is Gabrielle. I'm the Administrative Assistant at CReM. Leave us a short message down below. We will get back to you ASAP!

The Wilson lab is focused on two major aspects of regenerative medicine:

1) Developing gene therapy approaches for the study and treatment of lung diseases: The ability to manipulate gene expression in specified lung cell populations has both experimental and therapeutic potential for lung disease. By developing viral vectors that transduce specific lung cell types in vivo, we hope to minimize potential off-target effects while maximizing our ability to target diseased cell populations. We work with lentiviral and AAV vectors to overexpress or knockdown expression of genes important to disease pathogenesis in the lung.

2) Utilizing induced pluripotent stem cells (iPSC) to study human lung and liver diseases: The Wilson lab is interested in the application of patient-derived iPS cells for the study of lung and liver diseases, such as alpha-1 antitrypsin deficiency (AATD).

The Hawkins Lab is interested in how the human lung develops and responds to injury to better understand human lung disease. Induced pluripotent stem cells (iPSCs) offer a unique opportunity to model human lung disease and bridge the gap between research in animal models and humans.

Using this iPSC platform, we are focused on understanding the molecular mechanisms that control human lung development. We hope to apply this knowledge to advance our understanding of and develop precision medicine approaches for lung disease.

The Murphy laboratory is composed of dynamic and passionate researchers who utilize multiple stem cell-based platforms to answer basic biological questions and combat disease. Central directions of the laboratory include: developmental hematopoiesis, the modeling of blood-borne disease, and discovery and therapeutic intervention in sickle cell disease, amyloidosis, and aging.

The Murphy Lab has pioneered: The world’s largest sickle cell disease-specific iPSC library and platforms and protocols that can used to recapitulate hematopoietic ontogeny and to develop and validate novel therapeutic strategies for the disease; The successful modeling of a protein folding disorder called familial amyloidosis demonstrating the ability to model a long-term, complex, multisystem disease in a relatively short time, using lineage-specified cells (hepatic, cardiac and neuronal) derived from patient-specific stem cells; The first iPSC library created from subjects with exceptional longevity (centenarians) that serves as an unlimited resource of biomaterials to fuel the study of aging and the development of novel therapeutics for aging-related disease.

www.murphylaboratory.com

@DRGJMurphy

The Serrano Lab studies neurodevelopment and cardiovascular development in the context of rare multi-systemic disorders originated by pathogenic variants in epigenetic modifiers like KMT2D.  

We aim to identify shared molecular and cellular mechanisms driving cardiovascular and brain development with particular interest in cell differentiation, migration, and cell cycle progression.  

Our lab combines rare disease modeling in zebrafish together with cardiovascular and neurobiology techniques and human iPSC-derived brain organoids and endothelial cells.  

We believe that a patient-forward focus to our projects will help us to get better understanding of disease mechanisms through basic science research. To this end, we are active in the collaborative community among field experts and rare disease patient-advocacy groups who drive our research program to identify therapeutic targets in patient-specific iPS cells. 

The Mostoslavsky Lab is a basic science laboratory in the Section of Gastroenterology in the Department of Medicine at Boston University.

Our goal is to advance our understanding of stem cell biology with a focus on their genetic manipulation via gene transfer and their potential use for stem cell-based therapy.

The Mostoslavsky’s Lab designed and constructed the STEMCCA vector for the generation of iPS cells, a tool that has become the industry standard for nuclear reprogramming. Project areas in the lab focuses on the use of different stem cell populations, including embryonic stem cells, induced Pluripotent Stem (iPS) cells, hematopoietic stem cells and intestinal stem cells and their genetic manipulation by lentiviral vectors.

Our laboratory have already established a large library of disease-specific iPS cells with a particular interest in utilizing iPS cells to model diseases of the liver, the gastrointestinal tract, prion-mediated neurodegenerative diseases and immune-based inflammatory conditions, using iPSC-derived microglia, macrophages and T/NK cells.

The Gouon-Evans lab investigates cellular and molecular mechanisms driving liver development, regeneration and cancer. We specifically interrogate the role of progenitor/stem cells and how they share similar molecular signature and functions during these 3 processes.

Our innovative tools include: 1) directed differentiation of human pluripotent stem cells (PSC) to generate in vitro liver progenitors and their derivative hepatocytes, the main functional cell type of the liver, 2) mouse models with lineage tracing strategy to track in vivo the fate of progenitor cells, 3) PSC derivative cell transplantation into mouse models with damaged livers as cell therapy for liver diseases, 3) dissection of liver cancer specimens from patients to identify and define the impact of specific cancer stem cells in liver oncogenesis.

Projects in the Gouon-Evans lab will lead to a better understanding of the liver development, to the establishment of multi-modular approaches for improving liver regeneration with PSC derivatives, and will reveal the impact of specific cancer stem cells as a target for diagnosis and therapy in liver oncogenesis.