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Directed Differentiation Core Services

Directed Differentiation Core Services

January 2023/by Melissa Machado
PI: April Craft, PhD

Assistant Professor of Orthopedic Surgery, BCH, HMS; Affiliate Faculty, Stem Cells & Regenerative Biology; Principal Faculty, Harvard Stem Cell Institute; Co-Director, Musculoskeletal Disease Program, HSCI

Overview

The Craft lab develops and uses directed differentiation protocols for various skeletal and joint lineages from pluripotent stem cells (PSCs), including both embryonic stem cells and induced pluripotent stem cells, to study cell and tissue development and disease pathology.  Dr. Craft has been trained in the derivation of various mesoderm and endoderm lineages, with a focus on mesoderm-derived chondrocytes, articular and growth plate cartilage tissues, and cardiac lineages, and has new technologies emerging for tendon-ligament fibroblasts.  These existing protocols provide access to primitive streak mesoderm, endoderm, as well as paraxial mesoderm and mesenchymal-like progenitors.

Dr. Craft and her lab are available to discuss investigator-driven projects, experimental design, and provide hands-on training for generating working stocks of human iPSCs/ESCs and mouse PSCs, directed differentiations, deriving appropriate culture conditions and media compositions to support novel protocols, and troubleshooting existing protocols.  We also have experience with culturing and manipulating primary human skeletal cells (primarily chondrocytes and connective tissue lineages) and mouse embryonic limb bud micromass cultures. Our lab has purchased lot-tested reagents in bulk and we have a supportive infrastructure in place so that variability in PSC differentiations is minimized.

Example Services

  • Consultation with Dr. Craft (required)
    • 1 hour complementary to CSR members and trainees
  • Training or production in Craft lab
    • IPSC/ESC expansion and creation of master stock and working stocks (~50-100 vials total)
    • Human iPSC/ESC directed differentiation (full-length experiment)
      • Imaging: EVOS fluorescent microscope, Leica bright field microscope, Zeiss confocal
      • Flow cytometry available in BCH core via Craft lab
      • Other analyses such as RNA/histo at investigator’s expense
    • Mouse ESC/iPSC expansion and/or differentiation
    • Mouse limb bud micromass training (timed pregnant mice can be purchased if needed)
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Contact Us

April Craft
Email Address april.craft@childrens.harvard.edu

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