In vitro assessment of growth kinetics and differentiation potential of bone marrow derived mesenchymal and fetal fibroblast cells of caprine
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Keywords:
Differentiation, Fibroblast, Goat, Growth, MesenchymalAbstract
The present study assessed comparative growth kinetics and differentiation potential of mesenchymal and fetal fibroblast cells of goats in vitro. The mesenchymal cells were isolated from bone marrow and fibroblasts were obtained from skin of 3–4 months old fetus. During culture, more than 99% viability of both cell groups indicated optimum culture conditions for in vitro experiments. The population doubling time (PDT) of BM-MSCs and FFCs were similar at third (18.25±0.05 and 18.41±0.07) and seventh (19.61±0.06 and 19.68±0.12) passages. But at 12th passage, FFCs cells had a significantly higher PDT (35.77±0.68 vs 32.063±1.0) as compared to BM-MSCs. Both groups of cells showed a characteristic sigmoid curve on plotting cell concentration verses days of culture. The stem cell like property was exhibited by BM-MSCs by differentiating into osteoblast, chondrocytes and adipocytes under specific media, which were further confirmed by specific staining of cells. Also, the BM-MSCs showed colonogenic property by forming different number of clones at third (60.5±0.59), seventh (56.6±0.5) and 12th (33.4±0.45) passages. The fetal fibroblast cells did not exhibit colonogenic and differentiation potentials. This study concluded that although BM-MSCs and FFCs shared similar morphology and growth kinetics during initial passages but FFCs does not exhibit the stem cell like properties in terms of differentiation potency.
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Alt E, Yan Y, Gehmert S, Song Y H, Altman A, Gehmert S, Vykoukal D and Bai X. 2011. Fibroblasts share mesenchymal phenotypes with stem cells, but lack their differentiation and colony-forming potential. Biology of the Cell 103: 403. DOI: https://doi.org/10.1111/j.1768-322X.2011.tb01312.x
Alves F R, Guerra R R, Fioretto E T, Delgado J C, Machado- Júnior A A N, Ambrósio C E, Kerkis I and Miglino M A. 2010. Establishment of a protocol for obtention of neuronal stem cells lineages from the dog olfactory epithelium. Pesquisa Veterinária Brasileira 30 (4): 363–72. DOI: https://doi.org/10.1590/S0100-736X2010000400014
Augello A, Kurth T B and De Bari C. 2010. Mesenchymal stem cells: A perspective from in vitro cultures to in vivo migration and niches. European Cells and Materials 20: 121–33. DOI: https://doi.org/10.22203/eCM.v020a11
Blasi A, Martino C, Balducci L, Saldarelli M, Soleti A, Navone S E, Canzi L, Cristini S, Invernici G, Parati E A and Alessandri G. 2011. Dermal fibroblasts display similar phenotypic and differentiation capacity to fat-derived mesenchymal stem cells, but differ in anti-inflammatory and angiogenic potential. Vascular Cell 3: 5. DOI: https://doi.org/10.1186/2045-824X-3-5
Covas D T, Panepucci R A, Fontes A M, Silva W A Jr Orellana M D, Freitas M C, Neder L, Santos A R, Peres L C, Jamur M C and Zago M A. 2008. Multipotent mesenchymal stromal cells obtained from diverse human tissues share functional properties and gene-expression profile with CD146 perivascular cells and fibroblasts. Experimental Hematology 36: 642–54. DOI: https://doi.org/10.1016/j.exphem.2007.12.015
Cunningham B W, Sefter J C, Hu N and McAfee P C. 2010. Autologous growth factors versus autogenous graft for anterior cervical interbody fusion: an in vivo caprine model. Journal of Neurosurgery Spine 13 (2): 216–23. DOI: https://doi.org/10.3171/2010.3.SPINE09512
Da Rocha A R, Alves F R, Neto N M A, Dos Santos L F, De Almeida H M, De Carvalho Y K P, Bezerra D D O, Ferraz M S, Pessoa G T and De Carvalho M A M. 2012. Haematopoietic progenitor constituents and adherent cell progenitor morphology isolated from black-rumped agouti (Dasyprocta prymnolopha) bone marrow. Microscopy Research and Technique 75 (10): 1376–82. DOI: https://doi.org/10.1002/jemt.22077
Da Silva Meirelles L, Caplan A I and Nardi N B. 2008. In search of the in vivo identity of mesenchymal stem cells. Stem Cells 26: 2287–99. DOI: https://doi.org/10.1634/stemcells.2007-1122
Freshney R I. 2005. Culture of Animal Cells: A Manual of Basic Technique. Wiley-Liss, New York. DOI: https://doi.org/10.1002/0471747599.cac006
Friedenstein A J, Piatetzky-Shapiro II and Petrakova K V. 1966. Osteogenesis in transplants of bone marrow cells. Journal of Embryology and Experimental Morphology 16: 381–90. DOI: https://doi.org/10.1242/dev.16.3.381
Gotterbarm T, Breusch S J, Schneider U and Jung M. 2008. The minipig model for experimental chondral and osteochondral defect repair in tissue engineering: retrospective analysis of 180 defects. Laboratory Animals 42 (1): 71–82. DOI: https://doi.org/10.1258/la.2007.06029e
Haniffa M A, Collin M P, Buckley C D and Dazzi F. 2009. Mesenchymal stem cells: the fibroblasts’ new clothes. Haematologica 94: 258–63. DOI: https://doi.org/10.3324/haematol.13699
Jeong S W, Chu K, Jung K H, Kim S U, Kim M and Roh J K. 2003. Human neural stem cell transplantation promotes functional recovery in rats with experimental intracerebral hemorrhage. Stroke 34 (9): 2258–63. DOI: https://doi.org/10.1161/01.STR.0000083698.20199.1F
Kuroda Y, Kitada M, Wakaoa S, Nishikawa S, Makinoshima Y T H, Goda M, Akashi H, Inutsuka A, Niwa A, Shi-gemoto T, Nabeshima Y, Nakahata T, Nabeshima Y, Fujiyoshi Y and Dezawa M. 2010. Unique multipotent cells in adult human mesenchymal cell populations. Proceedings of the National Academy of Sciences of the United States of America 107: 8639–43. DOI: https://doi.org/10.1073/pnas.0911647107
Lechner F, Schütte A, Von-Bodungen U, Bertoni G, Pfister H, Jungi T W and Peterhans E. 1999. Inducible nitric oxide synthase is expressed in joints of goats in the late stage of infection with caprine arthritis encephalitis virus. Clinical and Experimental Immunology 117 (1): 70–75. DOI: https://doi.org/10.1046/j.1365-2249.1999.00932.x
Lorenz K, Sicker M, Schmelzer E, Rupf T, Salvetter J, Schulz- Siegmund M and Bader A. 2008. Multilineage differentiation potential of human dermal skin-derived fibroblasts. Experimental Dermatology 17: 925–32. DOI: https://doi.org/10.1111/j.1600-0625.2008.00724.x
Lysy P A, Smets F, Sibille C, Najimi M and Sokal E M. 2007. Human skin fibroblasts: from mesodermal to hepatocyte-like differentiation. Hepatology 46: 1574–85. DOI: https://doi.org/10.1002/hep.21839
Phinney D G and Prockop D J. 2007. Concise review: Mesenchymal stem/multipotent stromal cells: The state of transdifferentiation and modes of tissue repair—Current views. Stem Cells 25: 2896–902. DOI: https://doi.org/10.1634/stemcells.2007-0637
Sabatini F, Petecchia L, Tavian M, Jodon de Villeroche V, Rossi G A and Brouty-Boye D. 2005. Human bronchial fibroblasts exhibit a mesenchymal stem cell phenotype and multilineage differentiating potentialities. Laboratory Investigation 85: 962–71. DOI: https://doi.org/10.1038/labinvest.3700300
Sampaolesi M, Blot S, D’antona G, Granger N, Tonlorenzi R, Innocenzi A, Mognol P, Thibaud JL, Galvez B G, Barthélémy I, Perani L, Mantero S, Guttinger M, Pansarasa O, Rinaldi C, De Angelis MGC, Torrente Y, Bordignon C, Bottinelli R and Cossu G. 2006. Mesoangioblast stem cells ameliorate muscle function in dystrophic dogs. Nature 444 (7119): 574–79. DOI: https://doi.org/10.1038/nature05282
Vonk L A, Kroeze R J, Doulabi B Z, Hoogendoorn R J, Huang C, Helder M N, Everts V and Bank R A. 2010. Caprine articular, meniscus and intervertebral disc cartilage: An integral analysis of collagen network and chondrocytes. Matrix Biology 29 (3): 209–18. DOI: https://doi.org/10.1016/j.matbio.2009.12.001
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