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First Affiliated Hospital of Shenzhen University, Shenzhen Second People's Hospital

6 ARTICLES PUBLISHED IN JoVE

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Biology

Generation of Composite Plants in Medicago truncatula used for Nodulation Assays
Ying Deng *1, Guohong Mao *1, William Stutz 1, Oliver Yu 1
1Donald Danforth Plant Science Center

We demonstrate how hairy root composite plants can be used to study plant-rhizobium interactions and nodulation in the difficult-to-transform species Medicago truncatula.

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Biology

A Method for Islet Transplantation to the Omentum in Mouse
Ying Lu *1, Shangyou Zou *1, Suzanne Bertera *2, Rita Bottino 2, David K.C. Cooper 3, Zhengzhao Liu 1, Yi Huang 1, Chengjun Wang 1, Chungu Hong 1, Tian He 1, Hancheng Zhang 1, Qin Huo 4, Hongxing Fu 5, Zhiming Cai 1, Lisha Mou 1
1Shenzhen Xenotransplantation Medical Engineering Research and Development Center, Institute of Translational Medicine, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen University School of Medicine, Shenzhen University Health Science Center, 2Institute for Cellular Therapeutics, Allegheny-Singer Research Institute, 3Xenotransplantation Program/Department of Surgery, The University of Alabama at Birmingham, 4College of Life Science and Oceanography, Shenzhen University, 5School of Pharmcy, Wenzhou Medical College

A method for the omental transplantation of islets in a mouse is introduced. The isolated islets are mixed with hydrogel and the mixture is placed into the omental pouch of a diabetic mouse. Then, the blood glucose is monitored, and immuno-histochemical analysis is performed.

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Biology

Isolation and Culture of Primary Aortic Endothelial Cells from Miniature Pigs
Yanli Zhao 1,2,3, Chengjiang Zhao 4, David K.C. Cooper 5, Ying Lu 2, Kewang Luo 6, Huiyun Wang 3, Pengfei Chen 3, Changchun Zeng 3, Shaodong Luan 1, Lisha Mou 2, Hanchao Gao 1,2,3
1Department of Nephrology, Shenzhen Longhua District Central Hospital, Affiliated Central Hospital of Shenzhen Longhua District, Guangdong Medical University, 2Shenzhen Xenotransplantation Medical Engineering Research and Development Center, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen University School of Medicine, 3Department of Medical Laboratory, Shenzhen Longhua District Central Hospital, Affiliated Central Hospital of Shenzhen Longhua District, Guangdong Medical University, 4Department of Endocrinology, Shenzhen Longhua District Central Hospital, Affiliated Central Hospital of Shenzhen Longhua District, Guangdong Medical University, 5Xenotransplantation Program, Department of Surgery, University of Alabama at Birmingham, 6People's Hospital of Longhua

An effective enzymatic method for isolation of primary porcine aortic endothelial cells (pAECs) from miniature pigs is described. The isolated primary pAECs can be used to investigate the immune and coagulation response in xenotransplantation.

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Medicine

Inguinal Subcutaneous White Adipose Tissue (ISWAT) Transplantation Model of Murine Islets
Yuanzheng Peng *1, Zhicheng Zou *1, Jiao Chen 1, Hancheng Zhang 1, Ying Lu 1, Rito Bittino 2, Hongxing Fu 3, David K. C. Cooper 4, Shan Lin 1, Mengtao Cao 1, Yifan Dai 5, Zhiming Cai 1, Lisha Mou 1
1Shenzhen Xenotransplantation Medical Engineering Research and Development Center, Institute of Translational Medicine, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Health Science Center, 2Institute for Cellular Therapeutics, Allegheny-Singer Research Institute, 3School of Pharmaceutical Sciences, Wenzhou Medical University, 4Xenotransplantation Program/Department of Surgery, University of Alabama at Birmingham, 5Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University

In this protocol, a method of murine islet isolation and transplantation into the inguinal subcutaneous white adipose tissue is described. Isolated syngeneic murine islets are transplanted into a murine recipient using a basement membrane hydrogel. The blood glucose level of the recipients is monitored, and histology analysis of the islet grafts is performed.

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Biology

Quantification of Circulating Pig-Specific DNA in the Blood of a Xenotransplantation Model
Yangyang Deng *1,2, Ming Zhou *1,3, Ying Lu 1, Jiao Chen 1, Zuhui Pu 4, Dongjing Yu 1,5, Yifan Dai 6, Yongqiang Zhan 7, Lisha Mou 1
1Shenzhen Xenotransplantation Medical Engineering Research and Development Center, Institute of Translational Medicine, Shenzhen University Health Science Center, Shenzhen University School of Medicine, First Affiliated Hospital of Shenzhen University, Shenzhen Second People's Hospital, 2Xianning Hospital of Traditional Chinese Medicine, 3Liver-biotechnology (Shenzhen) Co., Ltd., 4Department of Radiology, Shenzhen University Health Science Center, Shenzhen University School of Medicine, First Affiliated Hospital of Shenzhen University, Shenzhen Second People's Hospital, 5Department of Life Sciences, University of Toronto, 6Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University, 7Department of Hepatopancreatobiliary Surgery, Shenzhen University Health Science Center, Shenzhen University School of Medicine, First Affiliated Hospital of Shenzhen University, Shenzhen Second People's Hospital

In this protocol, porcine specific primers were designed, plasmids-containing porcine specific DNA fragments were constructed, and standard curves for quantitation were established. Using species-specific primers, cpsDNA was quantified by qPCR in pig-to-mouse cell transplantation models and pig-to-monkey artery patch transplantation models.

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Biology

Adult Pig Islet Isolation
Ying Lu *1,2, Zuhui Pu *3, Jiao Chen 2, Jing Deng 2, Ying Deng 2,4, Shufang Zhu 2, Chunliang Xu 5, Fuwen Yao 1,2, Zijing Wu 1,2, Yong Ni 1, Yongqiang Zhan 1, Jun Cheng 1, Naiyang Zhan 1, Wenlong Huang 6, Zhiming Cai 2, Rita Bottino 7, Lisha Mou 2
1Department of Hepatopancreatobiliary Surgery, Shenzhen Institute of Translational Medicine, The First Affiliated Hospital of Shenzhen University, Shenzhen Second People's Hospital, 2Shenzhen Xenotransplantation Medical Engineering Research and Development Center, Shenzhen Institute of Translational Medicine, First Affiliated Hospital of Shenzhen University, Shenzhen Second People's Hospital, 3Imaging Department, Shenzhen Institute of Translational Medicine, The First Affiliated Hospital of Shenzhen University, Shenzhen Second People's Hospital, 4Faculty of Science, University of Waterloo, 5School of Life Science, The Chinese University of Hong Kong, 6General Surgery, The First Affiliated Hospital of Shantou University Medical College, 7Director Islet Programs, Imagine Pharma

This detailed protocol covers the methodological steps of adult pig islet isolation from the digestion phase via purification to the final functional assessment of the islets. This outline can be used as a guideline for adult pig islet isolation in research institutions.

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