30 September 2011
The culture of temporary tumor-like bone marrow mesenchymal stem cells (TT-BMSC) and the detection of cell biology property
Yi ChenABCDEF, Lin CongC, Ximeng YinE, Baotie DongD, Yaxin HanF, Guanjun TuADAnn Transplant 2011; 16(3): 49-58 :: ID: 881995
Abstract
Background: Bone marrow mesenchymal stem cells (BMSC) are common seed cells for transplantation. However, there are some limitations in their use that have not yet been resolved. Our research modified tumor-related genes temporarily on BMSC which simulated tumorigenesis temporarily in vitro. The cells were named temporarily tumor-like bone marrow mesenchymal stem cells (TT-BMSC).
Material/Methods: Cultivation of TT-BMSC: BMSC were cultured and identified, then the BMSC were transfected MMP-2 gene expressive vector and screened for 4 weeks by G418. Finally, the anti-oncogene PTEN of BMSC was knockdown by ribonucleic acid interference (RNAi) using PTEN gene special small interfering ribonucleic acid (SiRNA). The detection cell biology property of TT-BMSC in vitro: Methyl thiazolyl tetrazolium (MTT) assay and Cell cycle analysis for cell proliferation, Matrigel Invasion Assay for invasion and migration, and the cell model of ischemia and anoxia in vitro for survival.
Results: RT-PCR and Western blot results indicated MMP-2 expression increase significantly after transfection of the MMP-2 expressive vector in the BMSC, while PTEN mRNA and protein expression decrease significantly after PTEN RNAi, and the longest duration of the PTEN RNAi is 15 days. MTT assay and Cell cycle analysis indicated TT-BMSC cell growth vigor is reinforced significantly (P<0.001). Matrigel Invasion Assay showed that TT-BMSC can go through the matrigel successfully (P<0.001). The ability of TT-BMSC to tolerate ischemia and anoxia increased significantly in the model of ischemia and anoxia in vitro (P<0.001).
Conclusions: We cultivated TT-BMSC successfully and TT-BMSC possessed a powerful ability to survive, proliferate, invade and migrate in vitro.
Keywords: bone marrow mesenchymal stem cells, Matrix metalloproteinase-2, phosphatase and tensin homolog deleted on chromosome ten, ribonucleic acid interference
In Press
Original article
Living Donor Liver Transplantation with Small Left Lobe Grafts: Prospective Validation of Utility of Splene...Ann Transplant In Press; DOI: 10.12659/AOT.946374
Original article
A New Routine Immunity Score (RIS2020) to Predict Severe Infection in Solid-Organ Transplant RecipientsAnn Transplant In Press; DOI: 10.12659/AOT.946233
Original article
Survival Analysis of Liver Transplants in Patients with Acute Liver Failure from Acetaminophen and Mushroom...Ann Transplant In Press; DOI: 10.12659/AOT.946485
Original article
Medication Adherence Among Pediatric Post-Heart Transplant Patients in a Tertiary Care HospitalAnn Transplant In Press; DOI: 10.12659/AOT.946905
Most Viewed Current Articles
03 Jan 2023 : Original article 6,339
Impact of Autologous Stem Cell Transplantation on Primary Central Nervous System Lymphoma in First-Line and...DOI :10.12659/AOT.938467
Ann Transplant 2023; 28:e938467
16 May 2023 : Original article 6,002
Breaking Antimicrobial Resistance: High-Dose Amoxicillin with Clavulanic Acid for Urinary Tract Infections ...DOI :10.12659/AOT.939258
Ann Transplant 2023; 28:e939258
15 Aug 2023 : Review article 5,852
Free-Circulating Nucleic Acids as Biomarkers in Patients After Solid Organ TransplantationDOI :10.12659/AOT.939750
Ann Transplant 2023; 28:e939750
17 Jan 2023 : Original article 5,139
Non-Cryopreserved Peripheral Blood Stem Cell Graft for Autologous Hematopoietic Stem Cell Transplantation i...DOI :10.12659/AOT.938595
Ann Transplant 2023; 28:e938595