A novel approach for harvesting lymphatic endothelial cells from human foreskin dermis.
BACKGROUND
The lymphatic system plays an important role on tissue fluid and protein transportation, immune cell trafficking, lipid absorption, and even cancer metastasis, according to recent research. Disorders of the lymphatic system may lead to extremity edema and fibrosis. To investigate the physiological and pathophysiological characteristics of the lymphatic system, isolation and culture of lymphatic endothelial cells (LECs) in vitro are critical, although they remain difficult. The authors introduce a novel approach for harvesting LECs from child foreskin dermis.


RESULTS
After dispase digestion to remove epidermis, instead of the scrape procedure as described previously, collagenase was used to digest and harvest all the dermal cells, including vessel endothelial cells. Immunomagnetic beads were then applied to isolate CD34()/CD31(+)cells. These cells were characterized as LECs by immunofluorescent and RT-PCR with LECs specific markers including Prox-1, LYVE-1, and VEGFR-3. Proliferation of isolated cells in response to angiogenic factors and hypoxia were tested. Cells exhibited their typical cobblestone morphology as monolayer cultures under standard growth conditions and were propagated for at least seven passages without their characteristics being altered. LECs specifically responded to various stimuli, especially to VEGFs.


CONCLUSIONS
Using collagenase digestion procedure followed by immunomagnetic beads sorting, human dermal LECs could be harvested successfully.
Hu
Xueqing
X
Department of Plastic and Reconstructive Surgery, Shanghai 9th People's Hospital, Medical School of Shanghai Jiaotong University, Shanghai, China.
Jiang
Zhaohua
Z
Liu
Ningfei
N
D002478:Cells, Cultured; D002648:Child; D017364:Collagenases; D020405:Dermis / Q000166:cytology*; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D052816:Foreskin / Q000166:cytology*; D006801:Humans; D007150:Immunohistochemistry; D018189:Immunomagnetic Separation; D008297:Male; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017364:Collagenases