Improved Protocol to Study Osteoblast and Adipocyte Differentiation Balance.
| Title: | Improved Protocol to Study Osteoblast and Adipocyte Differentiation Balance. |
|---|---|
| Authors: | Alonso-Pérez A; Musculoskeletal Pathology Group, Institute IDIS, Santiago University Clinical Hospital, 15706 Santiago de Compostela, Spain.; Guillán-Fresco M; Musculoskeletal Pathology Group, Institute IDIS, Santiago University Clinical Hospital, 15706 Santiago de Compostela, Spain.; Franco-Trepat E; Musculoskeletal Pathology Group, Institute IDIS, Santiago University Clinical Hospital, 15706 Santiago de Compostela, Spain.; Jorge-Mora A; Musculoskeletal Pathology Group, Institute IDIS, Santiago University Clinical Hospital, 15706 Santiago de Compostela, Spain.; Traumatology and Orthopaedics Service, SERGAS, Santiago University Clinical Hospital, 15706 Santiago de Compostela, Spain.; López-Fagúndez M; Musculoskeletal Pathology Group, Institute IDIS, Santiago University Clinical Hospital, 15706 Santiago de Compostela, Spain.; Pazos-Pérez A; Musculoskeletal Pathology Group, Institute IDIS, Santiago University Clinical Hospital, 15706 Santiago de Compostela, Spain.; Crespo-Golmar A; Musculoskeletal Pathology Group, Institute IDIS, Santiago University Clinical Hospital, 15706 Santiago de Compostela, Spain.; Caeiro-Rey JR; Traumatology and Orthopaedics Service, SERGAS, Santiago University Clinical Hospital, 15706 Santiago de Compostela, Spain.; Gómez R; Musculoskeletal Pathology Group, Institute IDIS, Santiago University Clinical Hospital, 15706 Santiago de Compostela, Spain. |
| Source: | Biomedicines [Biomedicines] 2022 Dec 22; Vol. 11 (1). Date of Electronic Publication: 2022 Dec 22. |
| Publication Type: | Journal Article |
| Language: | English |
| Journal Info: | Publisher: MDPI AG Country of Publication: Switzerland NLM ID: 101691304 Publication Model: Electronic Cited Medium: Print ISSN: 2227-9059 (Print) Linking ISSN: 22279059 NLM ISO Abbreviation: Biomedicines Subsets: PubMed not MEDLINE |
| Imprint Name(s): | Original Publication: Basel, Switzerland : MDPI AG, [2013]- |
| Abstract: | Adipogenesis-osteoblastogenesis balance-rupture is relevant in multiple diseases. Current human mesenchymal stem cells (hMSCs) in vitro differentiation models are expensive, and are hardly reproducible. Their scarcity and variability make an affordable and reliable method to study adipocyte-osteoblast-equilibrium difficult. Moreover, media composition has been inconstant throughout the literature. Our aims were to compare improved differentiation lab-made media with consensus/commercial media, and to identify a cell-line to simultaneously evaluate both MSCs differentiations. Lab-made media were compared with consensus and commercial media in C3H10T1/2 and hMSC, respectively. Lab-made media were tested on aged women primary pre-osteoblast-like cells. To determine the optimum cell line, C3H10T1/2 and hMSC-TERT cells were differentiated to both cell fates. Differentiation processes were evaluated by adipocytic and osteoblastic gene-markers expression and staining. Lab-made media significantly increased consensus medium induction and overcame commercial media in hMSCs differentiation to adipocytes and osteoblasts. Pre-osteoblast-like cells only properly differentiate to adipocyte. Lab-made media promoted adipocyte gene-markers expression in C3H10T1/2 and hMSC-TERT, and osteoblast gene-markers in C3H10T1/2. Oil Red O and Alizarin Red staining supported these findings. Optimized lab-made media were better at differentiating MSCs compared to consensus/commercial media, and evidenced the adipogenic commitment of pre-osteoblast-like cells from aged-women. C3H10T1/2 is an optimum MSC line by which to study adipocyte-osteoblast differentiation balance. |
| References: | Osteoporos Int. 2018 Jul;29(7):1659-1664. (PMID: 29574516); Aging Cell. 2004 Dec;3(6):379-89. (PMID: 15569355); Stem Cells Dev. 2015 May 15;24(10):1150-63. (PMID: 25603196); Cell Physiol Biochem. 2013;31(2-3):452-61. (PMID: 23548631); J Mol Biol. 2008 Jun 6;379(3):428-42. (PMID: 18466921); Stem Cell Res Ther. 2018 Jul 27;9(1):203. (PMID: 30053888); Biochem Biophys Res Commun. 2005 Jan 21;326(3):527-38. (PMID: 15596132); BMC Dev Biol. 2010 May 07;10:47. (PMID: 20459638); Bone. 2007 Sep;41(3):462-73. (PMID: 17572166); Int J Mol Sci. 2021 Mar 10;22(6):. (PMID: 33802220); Eur Cell Mater. 2012 Jul 09;24:1-17. (PMID: 22777949); Osteoporos Int. 2012 Apr;23(4):1399-413. (PMID: 21881969); Coll Relat Res. 1981 Feb;1(2):209-26. (PMID: 7049548); J Biol Chem. 2016 Aug 19;291(34):17829-47. (PMID: 27402842); Exp Eye Res. 2013 Feb;107:65-73. (PMID: 23219871); Biochem Biophys Res Commun. 2018 Sep 5;503(2):715-721. (PMID: 29909008); Bone Res. 2015 Apr 14;3:15005. (PMID: 26273537); Cell Tissue Res. 1998 Nov;294(2):309-21. (PMID: 9799447); Annu Rev Biochem. 1995;64:345-73. (PMID: 7574486); Plast Reconstr Surg. 2010 Jul;126(1):41-52. (PMID: 20220555); Genes Dev. 2005 May 1;19(9):1093-104. (PMID: 15833911); Am J Pathol. 2014 Mar;184(3):697-713. (PMID: 24462663); Rheumatol Int. 2013 Sep;33(9):2255-60. (PMID: 23455632); Cell Tissue Bank. 2019 Jun;20(2):173-182. (PMID: 30887273); J Bone Miner Res. 2021 Apr;36(4):673-684. (PMID: 33314313); FASEB J. 2013 Jan;27(1):277-87. (PMID: 23047894); Methods Mol Biol. 2016;1416:149-58. (PMID: 27236670); J Bone Miner Res. 2003 Oct;18(10):1842-53. (PMID: 14584895); Bone. 2018 Jun;111:44-48. (PMID: 29551751); J Rheumatol. 2015 Nov;42(11):2055-8. (PMID: 26428208); Mol Cell. 1999 Oct;4(4):611-7. (PMID: 10549292); Exp Biol Med (Maywood). 2010 Oct;235(10):1185-93. (PMID: 20864461); Osteoporos Int. 2012 Feb;23(2):589-97. (PMID: 21359670); J Steroid Biochem Mol Biol. 2017 Jul;171:236-246. (PMID: 28408351); J Bone Miner Res. 1992 Feb;7(2):235-46. (PMID: 1373931); PLoS One. 2008 Feb 06;3(2):e1540. (PMID: 18253488); Nat Clin Pract Rheumatol. 2006 Jan;2(1):35-43. (PMID: 16932650); J Orthop Sci. 2004;9(5):503-8. (PMID: 15449126); Biogerontology. 2001;2(3):165-71. (PMID: 11708718); Ann Rheum Dis. 2013 Apr;72 Suppl 2:ii52-5. (PMID: 23253928); Proc Natl Acad Sci U S A. 1980 Jan;77(1):285-9. (PMID: 6928620); Nat Genet. 2019 Apr;51(4):716-727. (PMID: 30833796); Methods Enzymol. 2014;538:49-65. (PMID: 24529433); Calcif Tissue Int. 2012 Oct;91(4):286-96. (PMID: 22903508); Eur J Endocrinol. 2018 Oct 01;179(4):R165-R182. (PMID: 30299886); Nat Rev Rheumatol. 2009 Aug;5(8):442-7. (PMID: 19581903); Bone Miner. 1991 Jul;14(1):27-40. (PMID: 1868267); Proc Natl Acad Sci U S A. 2004 Jun 29;101(26):9607-11. (PMID: 15210946) |
| Grant Information: | PI16/01870 Instituto de Salud Carlos III; PI19/01446 Instituto de Salud Carlos III; Miguel Servet II Instituto de Salud Carlos III; Axudas predoutorais 2019 Instituto de Investigación Sanitaria de Santiago; MMA2018_2 Fundación Mutua Madrileña; CM20/00186 Instituto de Salud Carlos III; FI20/00210 Instituto de Salud Carlos III; MMA2020 Fundación Mutua Madrileña; FPU17/01706 Ministerio de Educación Cultura y Deporte |
| Contributed Indexing: | Keywords: adipocyte; differentiation; mesenchymal stem cell; osteoblast; osteoporosis; protocol |
| Entry Date(s): | Date Created: 20230121 Latest Revision: 20230123 |
| Update Code: | 20260130 |
| PubMed Central ID: | PMC9855576 |
| DOI: | 10.3390/biomedicines11010031 |
| PMID: | 36672539 |
| Database: | MEDLINE |
Journal Article