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The Fourth Military Medical University

6 ARTICLES PUBLISHED IN JoVE

Bioengineering

Sample Preparation Strategies for Mass Spectrometry Imaging of 3D Cell Culture Models

Dorothy R. Ahlf Wheatcraft^1,2, Xin Liu^1,2, Amanda B. Hummon^1,2

¹Department of Chemistry and Biochemistry, University of Notre Dame, ²Harper Cancer Research Institute, University of Notre Dame

Immortalized cancer cell lines can be grown as 3D cell cultures, a valuable model for biological research. This protocol describes mass spectrometry imaging of 3D cell cultures, including improvements in the sample preparation platform. The goal of this protocol is to instruct users to prepare 3D cell cultures for mass spectrometry imaging analysis.

Developmental Biology

Rapid Acquisition of 3D Images Using High-resolution Episcopic Microscopy

Haochuan Zhang^*1,2,3, JunGang Huang^*2,3,4, Xin Liu^2,3, Ping Zhu⁴, Zhongrong Li¹, Xue Li^2,3

¹Department of Pediatric Surgery, The Second Affiliated Hospital & Yuying Children's Hospital, Wenzhou Medical University, ²Departments of Urology, Boston Children's Hospital, ³Department of Surgery, Harvard Medical School, ⁴Department of Cardiovascular Surgery, Guangdong Cardiovascular Institute, Guangdong General Hospital, Guangdong Academy of Medical Sciences

We describe a detailed protocol using high-resolution episcopic microscopy to acquire three-dimensional (3D) images of mouse embryos. This improved protocol utilizes a modified tissue preparation method to enhance penetration of the fluorescent dye, thereby permitting morphometric analysis of both small and large-sized specimens.

Biology

Detecting Establishment of Shared Blood Supply in Parabiotic Mice by Caudal Vein Glucose Injection

Xin Liu¹, Xue Bai¹, Mingqi Li¹, Huimin Li¹, Yong Zhang^1,2, Baofeng Yang^1,3

¹Department of Pharmacology, The State-Province Key Laboratories of Biomedicine-Pharmaceutics of China, Key Laboratory of Cardiovascular Research, Ministry of Education, College of Pharmacy, Harbin Medical University, ²Institute of Metabolic Disease, Heilongjiang Academy of Medical Science, ³Department of Pharmacology and Therapeutics, Melbourne School of Biomedical Sciences, Faculty of Medicine, Dentistry and Health Sciences University of Melbourne

Here we describe a new method of detecting successful establishment of shared blood circulation of two parabionts through a caudal vein injection of glucose, which causes minimal damage and is not fatal to the parabionts.

Bioengineering

Isolation and Culture of Bone Marrow Mesenchymal Stem Cells from the Human Mandible

Chunlin Zong^*1, Lu Zhao^*1, Chong Huang², Yuanli Chen¹, Lei Tian¹

¹State Key Laboratory of Military Stomatology, National Clinical Research Center for Oral Diseases, Shaanxi Clinical Research Center for Oral Diseases, Department of Oral and Maxillofacial Surgery, School of Stomatology, The Fourth Military Medical University, ²College of Life Sciences, Northwest University

The present protocol describes an efficient procedure for isolating and culturing of human mandibular bone marrow-derived mesenchymal stem cells using the whole bone marrow adherence method. The cultured cells were identified by cell proliferation assays, flow cytometry, and multilineage differentiation induction.

Biology

Isolation and Analysis of Traceable and Functionalized Extracellular Vesicles from the Plasma and Solid Tissues

Yuan Cao^*1,2, Ji-Yu Qiu^*1,3, Da Chen^1,2, Chen-Yao Li^1,4, Shu-Juan Xing^1,5, Chen-Xi Zheng¹, Xin Liu², Yan Jin^1,6, Bing-Dong Sui¹

¹State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi International Joint Research Center for Oral Diseases, Center for Tissue Engineering, School of Stomatology, The Fourth Military Medical University, ²Department of Orthodontics, School of Stomatology, The Fourth Military Medical University, ³Department of VIP Dental Care, School of Stomatology, The Fourth Military Medical University, ⁴School of Basic Medicine, The Fourth Military Medical University, ⁵College of Life Science, Northwest University, ⁶Xi'an Institute of Tissue Engineering and Regenerative Medicine

The present protocol describes a method to extract extracellular vesicles from the peripheral blood and solid tissues with subsequent profiling of surface antigens and protein cargos.

Bioengineering

Construction, Characterization, and Regenerative Application of Self-Assembled Human Mesenchymal Stem Cell Aggregates

Shijie Li^1,2, Peisheng Liu^1,2,3, Ting Zhu^1,2,3, Bingdong Sui¹, Yan Jin¹, Kun Xuan², Chenxi Zheng¹

This study describes a method to construct aggregates based on the self-assembly of human mesenchymal stem cells and identifies the morphological and histological characteristics for the regenerative treatment of cranial bone defects.