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5 ARTICLES PUBLISHED IN JoVE

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Biology

Obtaining Hemocytes from the Hawaiian Bobtail Squid Euprymna scolopes and Observing their Adherence to Symbiotic and Non-Symbiotic Bacteria
Andrew J. Collins 1, Spencer V. Nyholm 1
1Department of Molecular and Cell Biology, University of Connecticut

This video will demonstrate how to obtain hemocytes (blood cells) from the Hawaiian bobtail squid, Euprymna scolopes for use in cell biological and bacterial adhesion assays. Hemocytes will be stained with a fluorescent dye and exposed to GFP-labeled bacteria.

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Medicine

Two Techniques to Create Hypoparathyroid Mice: Parathyroidectomy Using GFP Glands and Diphtheria-Toxin-Mediated Parathyroid Ablation
Ruiye Bi 1,2, Yi Fan 2,3, En Luo 2,4, Quan Yuan 2,4, Michael Mannstadt 1
1Endocrine Unit, Massachusetts General Hospital, Harvard Medical School, 2West China School of Stomatology, Sichuan University, 3Department of Oral Medicine, Infection and Immunity, Harvard School of Dental Medicine, 4State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University

Mice with acquired hypoparathyroidism would be useful for studying novel drug therapies for hypoparathyroidism. Two procedures to create such mice are demonstrated. The GFP-PTX mouse is generated by surgical parathyroidectomy guided by green fluorescing parathyroid glands. A second, non-surgical approach is based on parathyroid-specific expression of the diphtheria toxin receptor.

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Developmental Biology

Isolation, Culture, and Differentiation of Bone Marrow Stromal Cells and Osteoclast Progenitors from Mice
David E. Maridas 1, Elizabeth Rendina-Ruedy 1, Phuong T. Le 1, Clifford J. Rosen 1
1Maine Medical Center Research Institute

In this article, we present methods to isolate and differentiate bone marrow stromal cells and hematopoietic stem cells from mouse long bones. Two different protocols are presented yielding different cell populations suitable for expansion and differentiation into osteoblasts, adipocytes, and osteoclasts.

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Biology

Establishing Stable Binary Cultures of Symbiotic Saccharibacteria from the Oral Cavity
Andrew J. Collins *1,2, Pallavi P. Murugkar *1,2, Floyd E. Dewhirst 1,2
1The Forsyth Institute, 2Harvard School of Dental Medicine

We demonstrate a method for isolating difficult-to-grow members of the novel bacterial phylum, Saccharibacteria, by filtering dental plaque and co-culturing with host bacteria.

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Biology

Transverse Fracture of the Mouse Femur with Stabilizing Pin
Emily R. Moore 1, Marina Feigenson 2, David E. Maridas 1
1Department of Developmental Biology, Harvard School of Dental Medicine, 2Keros Therapeutics

This protocol describes a method to perform fractures on adult mice and monitor the healing process.

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