title: [The cultivation and identification of lacrimal gland adenoid cystic cancer stem cells].
abstract: OBJECTIVE
  To isolate and cultivate the Lacrimal gland Adenoid Cystic Carcinoma cells line, study Cancer Stem Cells properties.
  
  
  METHODS
  Experimental study. Lacrimal gland adenoid cystic carcinoma cancer stem cells were cultivated in serum-free suspension culture and the morphological changes were observed. Cells were divided into two groups, the LACC-CSC experimental group and the LACC control group. The flow cytometry instrument was used to detect the expression of classical stem cell markers CD133 and ABCG2. Transwell chamber was used to detect the cancer stem cell aggressivity and differentiated into the vascular endothelial cells. The tumorigenic force in vitro xenotransplantation were applied.
  
  
  RESULTS
  LACC cells can grow suspensively after vaccinated in serum free medium and form tumor microspheres after 10-12 days. Flow cytometry experiments showed that the expression ratio of stem cell markers CD133 in LACC-CSC was (35.67 ± 6.86)%, significantly different to LACC with (0.46 ± 0.48)%, (t = 8.867, P < 0.05). Similarly, the expression ratio of stem cell marker ABCG2 in LACC-CSC was (39.99 ± 4.54)%, significantly different to LACC with (6.75 ± 1.34)%, (t = -9.932, P < 0.05). In vitro experiment of Matrigel invasion, LACC-CSC went through the matrigel basement membrane averagely (32.60 ± 8.79)/HP contrary to LACC with average (10.20 ± 2.77)/HP after 24 hours, showing statistically significance (t = 5.433, P < 0.05) between the two groups. After training for 48 hours, the difference between two groups was still obvious (t = 5.779, P < 0.05) with LACC-CSC average (62.60 ± 4.83)/HP to LACC (44.00 ± 5.34)/HP. When induced by serum medium containing VEGF and bFGF, LACC-CSC grew adherent gradually and cell morphological changes occurred after continuous induction to long spindle cells. When cultured into three-dimensional matrix structure they formed vessel samples and expressed vascular endothelial marker CD31 and CD34. Transplanted tumor in vitro experiment, mice of LACC-CSC group grew tumors in 9 days with 100% tumorigenic rate, whereas LACC group 12 days with 100% tumorigenic rate.
  
  
  CONCLUSIONS
  LACC-CSC can be obtained through serum-free culture method. LACC-CSC grew suspensively and expressed classical stem cell markers. LACC-CSC were identified as cancer stem cells with stronger migration and invasion. LACC-CSC have tumorigenic force and multi-directional differentiation potential with general characteristics of the stem cell.
authors.affiliation: Tianjin Medical University Eye Hospital, Tianjin Medical University Eye Institute, School of Optometny and Ophthalmology, Tianjin 300384, China.
authors.forename: Jianmei
authors.initials: J
authors.lastname: Lyu
authors.affiliation: Email: yanjinhe88@gmail.com.
authors.forename: Yanjin
authors.initials: Y
authors.lastname: He
authors.forename: Lianfeng
authors.initials: L
authors.lastname: Xie
authors.forename: Xun
authors.initials: X
authors.lastname: Liu
authors.forename: Limin
authors.initials: L
authors.lastname: Zhu
mesh_terms: D000071916:AC133 Antigen; D000234:Adenoids / Q000473:pathology; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D003528:Carcinoma, Adenoid Cystic / Q000473:pathology*; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D016895:Culture Media, Serum-Free; D042783:Endothelial Cells / Q000473:pathology; D005134:Eye Neoplasms / Q000473:pathology*; D005434:Flow Cytometry; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D007765:Lacrimal Apparatus / Q000473:pathology; D007766:Lacrimal Apparatus Diseases / Q000473:pathology*; D051379:Mice; D014411:Neoplastic Stem Cells / Q000378:metabolism / Q000473:pathology*; D010455:Peptides / Q000378:metabolism
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D016895:Culture Media, Serum-Free; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse