[Establishment of a rat islets model coated with human umbilical vein endothelial cells coexpressing sCD40L-Ig and CTLA4-Ig].
OBJECTIVE
To isolate and culture primary human umbilical vein endothelial cells (HUVECs) and then to coat rat islets in vitro.


METHODS
Primary HUVECs were isolated and cultured, then they were identified by detecting human VIII factor associated antigen with immunohistochemical method. Weible-palade bodies were observed by transmission electron microscope and the phagocytosis for DiI-Ac-LDL was detected by fluorescent microscopy. Rat islets were isolated and coated with HUVECs coexpressing sCD40L-Ig and CTLA4-Ig in a heparinized culture dish. The coated islets were observed via morphology and high glucose challenge test.


RESULTS
The human VIII factor associated antigen, Weible-palade bodies and the phagocytosis for DiI-Ac-LDL were found in HUVECs. The islets coated by HUVECs coexpressing sCD40L-Ig and CTLA4-Ig were identified by green fluorescence proteins. Compared with that of the untreated islets (4.09), the release index of the coated islets was 3.94. There was no difference in insulin release between coated islets and untreated islets under glucose challenge.


CONCLUSIONS
HUVECs have been isolated, cultured and identified successfully. The rat islets were coated with HUVECs in vitro function well.
Li
Zhao-Lun
ZL
Department of Renal Transplantation, First Affiliated Hospital, Xi'an Jiaotong University, Xi'an 710061, China.
Xue
Wu-Jun
WJ
Tian
Pu-Xun
PX
Liu
Hua
H
Feng
Xin-Shun
XS
Ding
Xiao-Ming
XM
D000818:Animals; D015703:Antigens, CD / Q000276:immunology*; D023201:CD40 Ligand / Q000276:immunology*; D060908:CTLA-4 Antigen; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism* / Q000637:transplantation*; D005169:Factor VIII / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism*; D007150:Immunohistochemistry; D007515:Islets of Langerhans / Q000166:cytology*; D016381:Islets of Langerhans Transplantation; D008297:Male; D008856:Microscopy, Fluorescence; D023421:Models, Animal; D051381:Rats; D017207:Rats, Sprague-Dawley; D014471:Umbilical Veins / Q000166:cytology*
D015703:Antigens, CD; D060908:CTLA-4 Antigen; C556706:CTLA4 protein, human; C556708:Ctla4 protein, rat; D007074:Immunoglobulin G; D023201:CD40 Ligand; D005169:Factor VIII