University of California at Los Angeles

6 ARTICLES PUBLISHED IN JoVE

Biology

Microwave-assisted One-pot Synthesis of N-succinimidyl-4-[¹⁸F]fluorobenzoate ([¹⁸F]SFB)

Shuang Hou^*1,2,3, Duy Linh Phung^*1,2,3, Wei-Yu Lin^1,2,3, Ming-wei Wang⁴, Kan Liu⁵, Clifton Kwang-Fu Shen^1,2,3

¹Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California at Los Angeles, ²Crump Institute for Molecular Imaging, David Geffen School of Medicine, University of California at Los Angeles, ³California NanoSystems Institute, University of California at Los Angeles, ⁴Nuclear Medicine, PET Center, Shanghai Medical Collegea, Fudan University, ⁵Electronics and Information Engineering, College of Electronics and Information Engineering, Wuhan Textile University

A facile, one-pot synthesis of N-succinimidyl-4-[¹⁸F]fluorobenzoate ([¹⁸F]SFB) was developed based on a non-aqueous, three-step radiochemical process. Using microwave heating, the entire procedure can be completed in less than 30 min, or 60 min with further purification by preparative HPLC. The decay-corrected radiochemical yields (RCYs) were 35-5% (n > 30).

JoVE Journal

Use of Human Perivascular Stem Cells for Bone Regeneration

Aaron W. James^*1, Janette N. Zara^*2, Mirko Corselli², Michael Chiang¹, Wei Yuan², Virginia Nguyen¹, Asal Askarinam¹, Raghav Goyal¹, Ronald K. Siu³, Victoria Scott¹, Min Lee³, Kang Ting¹, Bruno Péault^2,4, Chia Soo²

¹Dental and Craniofacial Research Institute and Section of Orthodontics, School of Dentistry, UCLA, ²UCLA and Orthopaedic Hospital, Department of Orthopaedic Surgery and the Orthopaedic Hospital Research Center, UCLA, ³Department of Bioengineering, UCLA, ⁴Center for Cardiovascular Science, University of Edinburgh

Human perivascular stem cells (PSCs) are a novel stem cell class for skeletal tissue regeneration similar to mesenchymal stem cells (MSCs). PSCs can be isolated by FACS (fluorescence activated cell sorting) from adipose tissue procured during standard liposuction procedures, then combined with an osteoinductive scaffold to achieve bone formation in vivo.

Biology

Isolation of Blood-vessel-derived Multipotent Precursors from Human Skeletal Muscle

William C.W. Chen¹, Arman Saparov^2,3, Mirko Corselli⁴, Mihaela Crisan⁵, Bo Zheng⁶, Bruno Péault^7,8, Johnny Huard⁹

¹Stem Cell Research Center, Department of Bioengineering and Orthopedic Surgery, University of Pittsburgh, ²Department of Orthopedic Surgery, University of Pittsburgh, ³Nazarbayev University Research and Innovation System, Nazarbayev University, ⁴Department of Orthopaedic Surgery, UCLA Orthopaedic Hospital and the Orthopaedic Hospital Research Center, University of California at Los Angeles, ⁵Department of Cell Biology, Erasmus MC Stem Cell Institute, ⁶OHSU Center for Regenerative Medicine, Oregon Health & Science University, ⁷Centre for Cardiovascular Science and MRC Centre for Regenerative Medicine, Queen's Medical Research Institute and University of Edinburgh, ⁸David Geffen School of Medicine and the Orthopaedic Hospital Research Center, University of California at Los Angeles, ⁹Stem Cell Research Center, Department of Orthopedic Surgery and McGowan Institute for Regenerative Medicine, University of Pittsburgh

Blood vessels within human skeletal muscle harbor several multi-lineage precursor populations that are ideal for regenerative applications. This isolation method allows simultaneous purification of three multipotent precursor cell populations respectively from three structural layers of blood vessels: myogenic endothelial cells from intima, pericytes from media, and adventitial cells from adventitia.

Developmental Biology

Isolation of Perivascular Multipotent Precursor Cell Populations from Human Cardiac Tissue

James E. Baily¹, William C.W. Chen^2,3, Nusrat Khan⁴, Iain R. Murray⁴, Zaniah N. González Galofre⁴, Johnny Huard^5,6, Bruno Péault^4,7

¹Centre for Cardiovascular Science, Queen's Medical Research Institute, University of Edinburgh, ²Department of Bioengineering and Orthopaedic Surgery, University of Pittsburgh, ³Research Laboratory of Electronics and Department of Biological Engineering, Massachusetts Institute of Technology, ⁴MRC Centre for Regenerative Medicine, University of Edinburgh, ⁵Stem Cell Research Center, Department of Orthopaedic Surgery, University of Pittsburgh, ⁶Department of Orthopaedic Surgery, University of Texas Health Science Center at Houston, ⁷Department of Orthopaedic Surgery, UCLA Orthopaedic Hospital, David Geffen School of Medicine, University of California at Los Angeles

Human cardiac tissue harbours multipotent perivascular precursor cell populations that may be suitable for myocardial regeneration. The technique described here allows for the simultaneous isolation and purification of two multipotent stromal cell populations associated with native blood vessels, i.e. CD146⁺CD34^- pericytes and CD34⁺CD146^- adventitial cells, from the human myocardium.

Immunology and Infection

A Method for Targeted 16S Sequencing of Human Milk Samples

Nicole H. Tobin¹, Cora Woodward¹, Sara Zabih¹, David J. Lee¹, Fan Li¹, Grace M. Aldrovandi¹

¹Department of Pediatrics, University of California at Los Angeles

A semi-automated workflow is presented for targeted sequencing of 16S rRNA from human milk and other low-biomass sample types.

Biology

Human Adipose Tissue Micro-fragmentation for Cell Phenotyping and Secretome Characterization

Bianca Vezzani^1,2, Mario Gomez-Salazar¹, Joan Casamitjana¹, Carlo Tremolada³, Bruno Péault^1,4

¹MRC Center for Regenerative Medicine, University of Edinburgh, ²Dept. of Morphology, Surgery and Experimental Medicine, Section of General Pathology, University of Ferrara, ³Italian Image Institute, ⁴Orthopaedic Hospital Research Center and Broad Stem Cell Research Center, David Geffen School of Medicine, University of California

Here, we present human adipose tissue enzyme-free micro-fragmentation using a closed system device. This new method allows the obtainment of sub-millimeter clusters of adipose tissue suitable for in vivo transplantation, in vitro culture, and further cell isolation and characterization.