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Keratinocyte growth factor-2 and autologous serum potentiate the regenerative effect of mesenchymal stem cells in cornea damage in rats

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International Journal of Ophthalmology(English edition) 2014年第2期7卷 211-219页

作者： Ferda Alpaslan Pinarli Gülsen Okten Umit Beden Tunc Fisgin Mehmet Kefeli Nurten Kara Feride Duru Leman Tomak Department of Medical Biology and Genetics Faculty of Medicine Ondokuz Mayis University Department of Ophthalmology Faculty of MedicineOndokuz Mayis University Department of Pediatric Hematology Faculty of MedicineOndokuz Mayis University Department of Pathology Faculty of Medicine Ondokuz Mayis University Department of Medical Biostatistics Faculty of MedicineOndokuz Mayis University

AIM:To investigate the healing process after severe corneal epithelial damage in rats treated with mesenchymal stem cells(MSCs)cultured with or without keratinocyte growth factor(KGF-2)and autologous serum(AS)on amniotic membrane(AM).Many patients are blind and devastated by severe ocular surface diseases due to limbal stem cell *** marrow-derived MSCs are potential sources for cellbased tissue engineering to repair or replace the corneal tissue,having the potential to differentiate to epithelial ***:The study included 5 groups each including 10 female'Sprague Dawley'rats in addition to20 male rats used as bone marrow *** I rats received AM+MSCs,Group II rats AM+MSCs cultured with KGF-2,Group III rats AM+MSCs cultured with KGF-2+AS,Group IV rats only AM and Group V rats,*** was derived from blood drawn from male rats and bone marrow was obtained from the femur and tibia bones of the same *** effect was evaluated with clinical,histopathological and immunohistochemical *** engraftment was demonstrated via detection of donor genotype(Y+)in the recipient tissue(X)with polymerase chain ***:Corneal healing was significantly better in Groups I-III rats treated with MSC transplantation compared to Group IV and Group V rats with supportive treatment *** best results were obtained in Group III rats with 90%transparency,70%lack of neovascularization,and 100%epithelium damage limited to less than 1/4 of ***:We suggest that culture of MSCs with KGF-2 and AS on AM is effective in corneal repair in case of irreversible damage to limbal stem cells.

关键词： corneal wound healing mesenchymal stem cells keratinocyte growth factor-2 autologous serum amniotic membrane

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Effect of autologous serum after amniotic membrane transplantation for persistent corneal ulcers

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International Journal of Ophthalmology(English edition) 2024年第9期17卷 1639-1644页

作者： Rozaliya Hristova Petya Yankova Georgi Markov Alexander Oscar Yani Zdravkov Department of Ophthalmology Medical University SofiaUniversity Hospital AlexandrovskaSofia 1431Bulgaria Department of Clinical Immunology Medical University SofiaUniversity Hospital AlexandrovskaSofia 1431Bulgaria

AIM:To investigate the effect of adding autologous serum eye drops to the postoperative regime after amniotic membrane transplantation for severe persistent corneal ***:Forty eyes of 40 patients with persistent corneal ulcers were randomly assigned to artificial tears(sodium hyaluronate 0.2%,ATs group,n=20)or autologous serum eye drops(ASEDs,n=20)following treatment with amniotic membrane *** slit lamp images were acquired from all patients before and 30d post *** area with fibrovascular tissue was calculated using Image *** corneal sensitivity was assessed by Cochet-Bonnet aesthesiometry before and one month after *** tissue transparency was assessed with a novel optical ***:Mean age of patients was 61.65±16.47y and 57.3±19.11y in the ATs group and ASEDs group,***-two male and 18 female patients were included in the *** improvement in visual acuity was significantly greater in the ASEDs group(0.14±0.04)than the ATs(0.08±0.04;P=0.00046).Cochet-Bonnet aesthesiometry improved significantly after treatment with a similar rate between *** were no statistically significant differences in the area of postoperative fibrovascular tissue between the two groups(P=0.082).The success rate in the two groups was *** difference in densitometry between the ATs and ASEDs group was statistically significant(P=0.042)with greater reduction from baseline in the ASEDS ***:autologous serum eye drops can lead to better visual acuity,more stable results and improved densitometry and should be considered in the postoperative care following amniotic membrane transplantation.

关键词： autologous serum amniotic membrane ocular surface persistent corneal ulcer

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Effect of human autologous serum and fetal bovine serum on human corneal epithelial cell viability,migration and proliferation in vitro

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International Journal of Ophthalmology(English edition) 2017年第6期10卷 908-913页

作者： Ming-Feng Wu Tanja Stachon1 Berthold Seitz Achim Langenbucher Nora Szentmary Department of Ophthalmology Saarland University Medical Center Experimental Ophthalmology Saarland University Department of Ophthalmology Semmelweis University

AIM： To analyze the concentration-dependent effects of autologous serum （AS） and fetal bovine serum （FBS） on human corneal epithelial cell （HCEC） viability, migration and proliferation. METHODS： AS was prepared from 13 patients with non- healing epithelial defects Dulbecco＇s modified eagle medium/ Ham＇s F12 （DMEM/F12） with 5% FBS, 0.5% dimethyl sulphoxide （DMSO）, 10 ng/mL human epidermal growth factor, 1% insulin-transferrin-selenium, then were incubated in serum media： DMEM/F12 supplemented by 5%, 10%, 15% or 30% AS or FBS. HCEC viability was analyzed using cell proliferation kit XTI＇, migration using a wound healing assay, proliferation by the cell proliferation enzyme-linked immunosorbent assay （ELISA） BrdU kit. Statistical analysis was performed using the generalized linear model, the values at 30% AS or 30% FBS were used as the baselines. RESULTS： HCEC viability was the highest at 30% AS or 15% FBS and the lowest at 10% AS or 30% FBS application. HCEC migration was the quickest through 30% AS or 30% FBS and the slowest through 5% AS or 5% FBS concentrations. Proliferation was the most increased through 15% AS or 5% FBS and the least increased through 30% AS or 30% FBS concentrations. HCEC viability at 10% and 15% AS was significantly worse （P=0.001, P=0.023） compared to baseline and significantly better at 15% FBS （P=0.003） concentrations. HCEC migration was significantly worse （P〈0.007） and HCEC proliferation significantly better （P〈0.001） in all concentration groups compared to baseline. CONCLUSION： For the best viability of HCEC 30% AS or 15% FBS, for HCEC migration 30% AS or 30% FBS, for proliferation 15% AS or 5% FBS should be used. Therefore, we suggest the use of 30% AS in clinical practice.

关键词： autologous serum eye drops serum concen-tration migration proliferation viability human comeal epithelialcells

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autologous serum can induce mesenchymal stem cells into hepatocyte-like cells

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Journal of Medical Colleges of PLA(China) 2008年第2期23卷 63-69页

作者： Yang Yi Huo Jianhua Qu Bo Wu Shenli Zhang Mingyu Wang Zuoren Department of Hepatobiliary Surgery First Affiliated Hospital Medical College Xi ＂an Jiaotong University Xi＇an 710061 China Department of Heart First Affiliated Hospital Medical College Xi ＇an Jiaotong University Xi＇an 710061 China Department of Orthopaedics Second Affiliated Hospital Medical College Xi ＇an Jiaotong University Xi ＇an 710061 China

Objective： To investigate whether the rabbit serum after radiofrequency ablation to liver tumor can induce mesenchymal stem cells （MSCs） differentiating into hepatocyte-like cells in order to find a new source and culture process for repairing liver injury. Methods： A tumor piece of 1 mm× 1mm×1 mm was transplanted into a tunnel at right liver of rabbits. The model of liver tumor was established after 2-3 weeks. The serum was collected from rabbits 72 h after being subjected to radiofrequency ablation of the liver tumor. Mesenchymal stem cells were isolated from rabbit bone marrow and cultured in DMEM containing autologous rabbit serum. Three kinds of media （L-DMEM） were tested respectively： ① containing 10% fetal calf serum （FCS）; ② containing 30% rabbit autologous serum after radiofrequency ablation of the liver tumor （ASRF）; ③ containing 30% rabbit autologous serum （AS）. MSCs were cultured on 12-well plates until passage 2 and examined under the light and electron microscopy at indicted intervals. The expression of albumin and CKl8 was detected using immunofluorescence to identify the characteristics of differentiated cells. Results： MSCs performed differently in the presence of fetal calf serum, rabbit autologous serum and rabbit autologous serum after radiofrequency ablation of the liver tumor. Induced by the serum after radiofrequency ablation to liver tumor for 7 d, the spindle-shaped MSCs turned into round shaped and resembled hepatocyte-like cells. The reactions were not found in MSCs cultured in FCS and AS groups. After induction for 14 d, slender microvilli, cell-cell junction structure and cholangiole emerged, and the differentiated cells expressed albumin and CKl 8. All those could not been observed in 10% FCS and 30% autologous serum groups. Conclusion： Mesenchymal stem cells differentiate into hepatocyte-like cells in the serum after radiofrequency ablation of liver tumor, providing us a potential cell source and culture process for clinical applicatio

关键词： Radiofrequency ablation Tumor of liver autologous serum Mesenchymal stem cells Hepatocyte-like cells

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