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

6 ARTICLES PUBLISHED IN JoVE

Biology

Generation of Composite Plants in Medicago truncatula used for Nodulation Assays

Ying Deng^*1, Guohong Mao^*1, William Stutz¹, Oliver Yu¹

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.

Biology

A Method for Islet Transplantation to the Omentum in Mouse

Ying Lu^*1, Shangyou Zou^*1, Suzanne Bertera^*2, Rita Bottino², David K.C. Cooper³, Zhengzhao Liu¹, Yi Huang¹, Chengjun Wang¹, Chungu Hong¹, Tian He¹, Hancheng Zhang¹, Qin Huo⁴, Hongxing Fu⁵, Zhiming Cai¹, Lisha Mou¹

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.

Biology

Isolation and Culture of Primary Aortic Endothelial Cells from Miniature Pigs

Yanli Zhao^1,2,3, Chengjiang Zhao⁴, David K.C. Cooper⁵, Ying Lu², Kewang Luo⁶, Huiyun Wang³, Pengfei Chen³, Changchun Zeng³, Shaodong Luan¹, Lisha Mou², 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.

Medicine

Inguinal Subcutaneous White Adipose Tissue (ISWAT) Transplantation Model of Murine Islets

Yuanzheng Peng^*1, Zhicheng Zou^*1, Jiao Chen¹, Hancheng Zhang¹, Ying Lu¹, Rito Bittino², Hongxing Fu³, David K. C. Cooper⁴, Shan Lin¹, Mengtao Cao¹, Yifan Dai⁵, Zhiming Cai¹, Lisha Mou¹

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.

Biology

Quantification of Circulating Pig-Specific DNA in the Blood of a Xenotransplantation Model

Yangyang Deng^*1,2, Ming Zhou^*1,3, Ying Lu¹, Jiao Chen¹, Zuhui Pu⁴, Dongjing Yu^1,5, Yifan Dai⁶, Yongqiang Zhan⁷, Lisha Mou¹

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.

Biology

Adult Pig Islet Isolation

Ying Lu^*1,2, Zuhui Pu^*3, Jiao Chen², Jing Deng², Ying Deng^2,4, Shufang Zhu², Chunliang Xu⁵, Fuwen Yao^1,2, Zijing Wu^1,2, Yong Ni¹, Yongqiang Zhan¹, Jun Cheng¹, Naiyang Zhan¹, Wenlong Huang⁶, Zhiming Cai², Rita Bottino⁷, Lisha Mou²

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.