title: [Study on oriented differentiation of bone marrow mesenchymal stem cells by fibroblast in rat uterine ligament with mechanical stretch].
abstract: OBJECTIVE
  To investigate the effect on the differentiation of bone marrow mesenchymal stem cells (BMSC) with non-contact co-culture with mechanical stimulated ligament fibroblasts.
  
  
  METHODS
  A cyclic 10% uniaxia strain at 1 Hz was applied on rat pelvic ligament fibroblasts, then were co-cultured with BMSC for 3, 6 and 12 days in non-contact condition. The protein expression of collagen I, III in BMSC were detected by SP method and revealed by the mean gray value. The mRNA expressions of collagens type I and type III in the BMSCs were measured with real-time (RT)-PCR, and the results were indicated by the ratio between the mRNA and glyceraldehyde-3-phosphate dehydrogenase (GAPDH).
  
  
  RESULTS
  (1) Protein expression: after 3 days co-culture with pelvic ligament fibroblasts, expression of collagen I and III in BMSC are 82.4±3.4 and 76.8±2.5. When compared with 80.2±2.6 and 74.6±1.1 in BMSC without co-culture, there was no significant difference (P>0.05). After 6 days co-culture with pelvic ligament fibroblasts, the expression of collagen I and III of 126.6±2.2 and 118.6±1.4 in BMSC were significantly higher than 82.7±3.0 and 76.2±1.3 in BMSC without co-culture (P<0.05). Similarly, after 12 days co-culture with pelvic ligament fibroblasts, the expression of collagen I and III of 135.3±3.4 and 128.7±2.6 in BMSC were significantly higher than 86.6±1.3 and 81.8±1.4 in BMSC without co-culture (P<0.05). (2) mRNA expression: after 3 days co-culture with pelvic ligament fibroblasts, the mRNA expression of type I and type III collagens in BMSC are 2.10±0.20 and 1.20±0.30. When compared with mRNA expression of 2.01±0.12 and 1.13±0.21 in BMSC without co-culture, no significant difference were observed (P>0.05). After 6 days co-culture with pelvic ligament fibroblasts, the mRNA expressions of type I and type III collagens mRNA were 5.60±0.21 and 2.61±0.20, which were significantly higher than 3.70±0.33 and 1.82±0.14 in BMSC without coculture (P<0.05). After 12 days co-culture with pelvic ligament fibroblasts, the mRNA expressions of type I and type III collagens of 5.91±0.31 and 2.92±0.23 were significantly higher than 4.04±0.21 and 2.04±0.13 in BMSC without co-culture (P<0.05).
  
  
  CONCLUSIONS
  Non-contact co-culture with mechanical stretch stimulated ligament fibroblasts, it might promote synthesis of types Iand III collagen in rat BMSCs and induced BMSC differentiated into pelvic ligament fibroblasts.
authors: Department of Obstetrics and Gynecology, Third Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
authors: Chen-chen
authors: CC
authors: Ren
authors: Rui-fang
authors: RF
authors: Ren
authors: Bing
authors: B
authors: Zhao
authors: Xi
authors: X
authors: Zhang
authors: Yong-jie
authors: YJ
authors: Jiang
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology; D002454:Cell Differentiation; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D024061:Collagen Type III / Q000235:genetics / Q000378:metabolism; D005260:Female; D005347:Fibroblasts / Q000166:cytology* / Q000648:ultrastructure; D008022:Ligaments / Q000166:cytology*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction; D013314:Stress, Mechanical; D014599:Uterus
chemical_list: D024042:Collagen Type I; D024061:Collagen Type III; D012333:RNA, Messenger