Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The ONWARDS insulin icodec trials

A round-up of the ONWARDS phase 3 clinical trials evaluating the novel weekly insulin icodec in people with diabetes.
ADVANCED SEARCH
Log in
Top
Comparative Clinical Pathology
Published in: Comparative Clinical Pathology 6/2015

01-11-2015 | Original Article

Genetic evaluation of bone marrow-derived mesenchymal stem cells by a modified karyotyping method

Authors: Seyedeh-Leili Asadi-Yousefabad, Azizollah Khodakaram-Tafti, Mehdi Dianatpour, Davood Mehrabani, Shahrokh Zare, Amin Tamadon, Saman Nikeghbalian, Alireza Raayat-Jahromi, Somayeh Ahmadlou

Published in: Comparative Clinical Pathology | Issue 6/2015

Login to get access

Abstract

Bone marrow is the most important source of mesenchymal stem cells (MSCs). Significant development in the comprehension of MSCs biology has opened the way to their clinical use. The object of this study was to establish a method for karyotyping of canine MSCs. We examined the effect of the increase in passage number on the chromosome and appearance of MSCs. A large 3-year-old mixed breed dog was used. Bone marrow (20 ml) was collected from the proximal humerus under general anesthesia and cultured and passaged in alpha minimal essential medium (αMEM) and fetal bovine serum (FBS). The canine bone marrow MSCs (BM-MSCs) were characterized by reverse transcription polymerase chain reaction (RT-PCR) and flask adherent fibroblast-like morphology. Karyotyping of cells of passages 1 and 8 was performed. The chromosome numbers were counted and evaluated to obtain the percentage of diploid cells. The best time for exposing BM-MSCs with media containing colcemid was 10 h. These cells were positive for the BM-MSCs markers CD90 and negative for hematopoietic markers CD34 and CD45 by RT-PCR. Karyotyping of these cells was normal, and also fibroblast-like cells were observed in every passage.
Literature
Berman D, Willman M, Han D, Kleiner G, Kenyon N, Cabrera O, Karl J, Wiseman R, O’Connor D, Bartholomew A (2010) Mesenchymal stem cells enhance allogeneic islet engraftment in nonhuman primates. Diabetes 59:2558–2568PubMedCentralCrossRefPubMed
Csaki C, Matis U, Mobasheri A, Ye H, Shakibaei M (2007) Chondrogenesis, osteogenesis and adipogenesis of canine mesenchymal stem cells: a biochemical, morphological and ultrastructural study. Histochem Cell Biol 128:507–520CrossRefPubMed
Dissanayaka W, Zhu XM, Zhang C, Jin L (2011) Characterization of dental pulp stem cells isolated from canine premolars. J Endod 37:1074–1080CrossRefPubMed
Heim S, Mitelman F (2011) Cancer cytogenetics: chromosomal and molecular genetic abberations of tumor cells. Wiley pp 65–70
Izadpanah R, Kaushal D, Kriedt C, Tsien F, Patel B, Dufour J, Bunnell B (2008) Long-term in vitro expansion alters the biology of adult mesenchymal stem cells. Cancer Res 68:4229–4238PubMedCentralCrossRefPubMed
Jafarian M, Eslaminejad-Baghaban M, Khojasteh A, Mashhadi-Abbas F, Dehghan M, Hassanizadeh R, Houshmand B (2008) Marrow-derived mesenchymal stem cells-directed bone regeneration in the dog mandible: a comparison between biphasic calcium phosphate and natural bone mineral. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 105:14–24CrossRef
Jin H, Bae Y, Kim M, Kwon S, Jeon H, Choi S, Kim S, Yang Y, Oh W, Chang J (2013) Comparative analysis of human mesenchymal stem cells from bone marrow, adipose tissue, and umbilical cord blood as sources of cell therapy. Int J Mol Sci 14:17986–18001PubMedCentralCrossRefPubMed
Jones E, Kinsey S, English A, Jones R, Straszynski L, Meredith D, Markham A, Jack A, Emery P, McGonagle D (2002) Isolation and characterization of bone marrow multipotential mesenchymal progenitor cells. Arthritis Rheum 46:3349–3360CrossRefPubMed
Kamishina H, Deng J, Oji T, Cheeseman J, Clemmons R (2006) Expression of neural markers on bone marrow–derived canine mesenchymal stem cells. Am J Vet Res 67:1921–1928CrossRefPubMed
Kim S, Im K, Park S, Kwon J, Kim J, Choi Q, Hwang S, Han S, Kwon S, Oh I (2014) Asymmetric aneuploidy in mesenchymal stromal cells detected by in situ karyotyping and fluorescence in situ hybridization: suggestions for reference values for stem cells. Stem Cells Dev 24(1):77–92CrossRef
King R, Mulligan P, Stansfield W (2012) A dictionary of genetics. Oxford University Press, pp 242
Langford C, Fischer P, Binns M, Holmes N, Carter N (1996) Chromosome-specific paints from a high-resolution flow karyotype of the dog. Chromosome Res 4:115–123CrossRefPubMed
Mareschi K, Ferrero I, Rustichelli D, Aschero S, Gammaitoni L, Aglietta M, Madon E, Fagioli F (2006) Expansion of mesenchymal stem cells isolated from pediatric and adult donor bone marrow. J Cell Biochem 97:744–754CrossRefPubMed
Mehrabani D, Mahboobi R, Dianatpour M, Zare Sh, Tamadon A, Hosseini SE (2014a) Establishment, culture and characterization of Guinea pig fetal fibroblast cells. Vet Med Int Article ID 510328, 5 pages
Mehrabani D, Rahmanifar F, Mellinejad M, Tamadon A, Dianatpour M, Zare S, Razeghian Jahromi I, Ghobadi F (2014b) Isolation, culture, characterization, and adipogenic differentiation of heifer endometrial mesenchymal stem cells. Comp Clin Pathol. doi:10.​1007/​s00580-014-2053-0
Neagu M, Suciu E, Ordodi V, Paunescu V (2005) Human mesenchymal stem cells as basic tools for tissue engineering: isolation and culture. Rom J Biophys 15:29–34
Neupane M, Chang C, Kiupel M, Yuzbasiyan-Gurkan V (2008) Isolation and characterization of canine adipose-derived mesenchymal stem cells. Tissue Eng Part A 14:1007–1015CrossRefPubMed
Patterson D (2000) Companion animal medicine in the age of medical genetics. J Vet Intern Med 14:1–9CrossRefPubMed
Perry B, Zhou D, Wu X, Yang Byers M, Chu T, Hockema J, Woods E, Goebel W (2008) Collection, cryopreservation, and characterization of human dental pulp-derived mesenchymal stem cells for banking and clinical use. Tissue Eng Part C Methods 14:149–156PubMedCentralCrossRefPubMed
Ribeiro J, Pereira T, Amorim I, Caseiro A, Lopes M, Lima J, Gartner A, Santos J, Bartolo P, Rodrigues J (2014) Cell therapy with human MSCs isolated from the umbilical cord Wharton jelly associated to a PVA membrane in the treatment of chronic skin wounds. Int J Med Sci 11:979–987PubMedCentralCrossRefPubMed
Semenov O, Koestenbauer S, Riegel M, Zech N, Zimmermann R, Zisch A, Malek A (2010) Multipotent mesenchymal stem cells from human placenta: critical parameters for isolation and maintenance of stemness after isolation. Am J Obstet Gynecol 202:113–191
Smith M (2013) Isolation and characterization of stem cells literatures. Stem Cells 4:73–109
Sreekumar T, Ansari M, Chandra G, Sharma T (2014) Isolation and characterization of Buffalo Wharton’s Jelly derived mesenchymal stem cells. J Stem Cell Res Ther 4:1–6
Switonski M, Rogalska-Niznik N, Szczerbal I, Baer M (2003) Review: chromosome polymorphism and karyotype evolution of four canids: the dog, red fox, arctic fox and raccoon dog. Caryologia 56:375–385CrossRef
Tharasanit T, Phutikanit N, Wangdee C, Soontornvipart K, Tantrajak S, Kaewamatawong T, Suwimonteerabutr J, Supaphol P, Techakumphu M (2011) Differentiation potentials of canine bone marrow mesenchymal stem cells. Thai J Vet Med 41:79–86
Tien J, Lau L, Tien S, Tan M (2014) The clinical utility of conventional karyotyping in the detection of cytogenetic abnormalities in soft tissue tumours: an Asian institutional experience. Hong Kong Med J 20:1–8
Torkzaban P, Saffarpour A, Bidgoli M, Sohilifar S (2012) In vitro evaluation of isolation possibility of stem cells from intra oral soft tissue and comparison of them with bone marrow stem cells. J Dent 9:1–9
Vieira N, Brandalise V, Zucconi E, Secco M, Strauss B, Zatz M (2010) Isolation, characterization, and differentiation potential of canine adipose-derived stem cells. Cell Transplant 19:279–289CrossRefPubMed
Zhang H, Chen Z, Bie P (2012) Bone marrow–derived mesenchymal stem cells as immunosuppressants in liver transplantation: a review of current data. Transfus Med Rev 26:129–141CrossRefPubMed
Zucconi E, Vieira N, Bueno D, Secco M, Jazedje T, Ambrosio C, Passos-Bueno M, Miglino M, Zatz M (2010) Mesenchymal stem cells derived from canine umbilical cord vein—a novel source for cell therapy studies. Stem Cells Dev 19:395–402CrossRefPubMed
Metadata
Title
Genetic evaluation of bone marrow-derived mesenchymal stem cells by a modified karyotyping method
Authors
Seyedeh-Leili Asadi-Yousefabad
Azizollah Khodakaram-Tafti
Mehdi Dianatpour
Davood Mehrabani
Shahrokh Zare
Amin Tamadon
Saman Nikeghbalian
Alireza Raayat-Jahromi
Somayeh Ahmadlou
Publication date
01-11-2015
Publisher
Springer London
Published in
Comparative Clinical Pathology / Issue 6/2015
Print ISSN: 1618-5641
Electronic ISSN: 1618-565X
DOI
https://doi.org/10.1007/s00580-015-2081-4

Other articles of this Issue 6/2015

Comparative Clinical Pathology 6/2015 Go to the issue

Original Article

Immunoexpression of metalloproteinase 14 and tissue inhibitor of metalloproteinase 2 in colorectal carcinomas and lymph node metastases

Original Article

The relationship between bovine adenomyosis and endometritis

Original Article

Serum protein profile of hookworm infection in dogs

Original Article

Sub-acute toxicity of lantadenes isolated from Lantana camara leaves in guinea pig animal model

Original Article

Hereditary xanthinuria and urolithiasis in a domestic shorthair cat

Original Article

Antidiabetic effect of Pistacia atlantica and Amygdalus scoparia in streptozotocin-induced diabetic mice