Accedi

Boston University and Boston Medical Center

3 ARTICLES PUBLISHED IN JoVE

Biology

Generation of Human Induced Pluripotent Stem Cells from Peripheral Blood Using the STEMCCA Lentiviral Vector

Andreia Gianotti Sommer¹, Sarah S. Rozelle¹, Spencer Sullivan², Jason A. Mills³, Seon-Mi Park¹, Brenden W. Smith¹, Amulya M. Iyer¹, Deborah L. French³, Darrell N. Kotton¹, Paul Gadue³, George J. Murphy¹, Gustavo Mostoslavsky¹

¹Center for Regenerative Medicine (CReM), Boston University School of Medicine, ²Department of Hematology, Children's Hospital of Philadelphia, ³Center for Cellular and Molecular Therapeutics, Children's Hospital of Philadelphia

Here we show a simple and effective protocol for the generation of human iPSCs from 3-4 ml of peripheral blood using a single lentiviral reprogramming vector. Reprogramming of readily available blood cells promises to accelerate the utilization of iPSC technology by making it accessible to a broader research community.

Developmental Biology

Generating 3D Spheres and 2D Air-Liquid Interface Cultures of Human Induced Pluripotent Stem Cell-Derived Type 2 Alveolar Epithelial Cells

Rhiannon B. Werder^*1,2,3, Jessie Huang^*1,2, Kristine M. Abo^1,2, Olivia T. Hix^1,2, Kasey Minakin^1,2, Konstantinos-Dionysios Alysandratos^1,2, Carly Merritt^1,2, Kayleigh Berthiaume^1,2, Andrea B. Alber^1,2, Claire L. Burgess^1,2, Darrell N. Kotton^*1,2, Andrew A. Wilson^*1,2

¹Center for Regenerative Medicine, Boston University and Boston Medical Center, ²The Pulmonary Center and Department of Medicine, Boston University School of Medicine, ³QIMR Berghofer Medical Research Institute

The present protocol describes human induced pluripotent stem cell-derived type 2 alveolar epithelial-like cells (iAT2s). These cells can be cultured as self-renewing spheres in 3D culture or adapted to air-liquid interface (ALI) culture.

Developmental Biology

Generation of Airway Epithelial Cell Air-Liquid Interface Cultures from Human Pluripotent Stem Cells

Andrew Berical^1,2, Mary Lou Beermann², Shingo Suzuki³, Jake LeSuer², Taylor Matte², Brian Davis³, Darrell Kotton^1,2, Finn Hawkins^1,2

¹Pulmonary Center, Boston University School of Medicine, ²Center for Regenerative Medicine, Boston University and Boston Medical Center, ³Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center

Recent advances in human induced pluripotent stem cell differentiation protocols allow for the stepwise derivation of organ-specific cell types. Here, we provide detailed steps for the maintenance and expansion of iPSC-derived airway basal cells and their differentiation into a mucociliary epithelium in air-liquid interface cultures.