التفاصيل البيبلوغرافية
العنوان:
Immortalization and characterization of osteoblast cell lines generated from wild-type and Nmp4-null mouse bone marrow stromal cells using murine telomerase reverse transcriptase (mTERT)
المؤلفون:
Rita Gerard-O'Riley , Marta Alvarez , Joseph P. Bidwell , Brittney-Shea Herbert , Alexander G. Robling , Binu K. Philip , Michael Hanlon , Paul Childress , Fredrick M. Pavalko
المصدر:
Journal of cellular physiology . 227(5)
سنة النشر:
2011
مصطلحات موضوعية:
Telomerase , medicine.medical_specialty , Stromal cell , Physiology , Clinical Biochemistry , Parathyroid hormone , Bone Morphogenetic Protein 2 , Bone Marrow Cells , Biology , Bone morphogenetic protein 2 , Article , Cell Line , Mice , Nuclear Matrix-Associated Proteins , Internal medicine , medicine , Adipocytes , Animals , Humans , Telomerase reverse transcriptase , Mice, Knockout , Osteoblasts , Osteoblast , Cell Biology , Molecular biology , BMPR1A , Mice, Inbred C57BL , medicine.anatomical_structure , Endocrinology , Phenotype , Parathyroid Hormone , Female , Bone marrow , Stromal Cells , Transcription Factors
الوصف:
Intermittent parathyroid hormone (PTH) adds new bone to the osteoporotic skeleton; the transcription factor Nmp4/CIZ represses PTH-induced bone formation in mice and as a consequence is a potential drug target for improving hormone clinical efficacy. To explore the impact of Nmp4/CIZ on osteoblast phenotype, we immortalized bone marrow stromal cells from wildtype (WT) and Nmp4-knockout (KO) mice using murine telomerase reverse transcriptase. Clonal lines were initially chosen based on their positive staining for alkaline phosphatase and capacity for mineralization. Disabling Nmp4/CIZ had no gross impact on osteoblast phenotype development. WT and KO clones exhibited identical sustained growth, reduced population doubling times, extended maintenance of the mature osteoblast phenotype, and competency for differentiating toward the osteoblast and adipocyte lineages. Additional screening of the immortalized cells for PTH-responsiveness permitted further studies with single WT and KO clones. We recently demonstrated that PTH-induced c-fos femoral mRNA expression is enhanced in Nmp4-KO mice and in the present study we observed that hormone stimulated either an equivalent or modestly enhanced increase in c-fos mRNA expression in both primary null and KO clone cells depending on PTH concentration. The null primary osteoblasts and KO clone cells exhibited a transiently enhanced response to bone morphogenetic protein 2 (BMP2). The clones exhibited lower and higher expressions of the PTH receptor (Pthr1) and the BMP2 receptor (Bmpr1a, Alk3), respectively, as compared to primary cells. These immortalized cell lines will provide a valuable tool for disentangling the complex functional roles underlying Nmp4/CIZ regulation of bone anabolism. J. Cell. Physiol. 227: 1873–1882, 2012. © 2011 Wiley Periodicals, Inc.
تدمد:
1097-4652
URL الوصول:
https://explore.openaire.eu/search/publication?articleId=doi_dedup___::b4e453e1280cbd40fb8a8dd8df1a4de2 https://pubmed.ncbi.nlm.nih.gov/21732358
حقوق:
OPEN
رقم الأكسشن:
edsair.doi.dedup.....b4e453e1280cbd40fb8a8dd8df1a4de2
قاعدة البيانات:
OpenAIRE
