Title: Viability and expression pattern of cryopreserved mesenchymal stem cells derived from buffalo bone marrow
Authors: Pooja Devi, Manjinder Sharma, DV Singh, MK Lonare and Rahul Udehiya
Source: Ruminant Science (2017)-6(1):7-12.
Cite this reference as: Devi Pooja, Sharma Manjinder, Singh DV, Lonare MK and Udehiya Rahul (2017).Viability and expression pattern of cryopreserved mesenchymal stem cells derived from buffalo bone marrow. Ruminant Science 6(1):7-12.
The present investigation was carried out to compare the viability and expression pattern of cryopreserved mesenchymal stem cells (MSCs) with fresh MSCs derived from buffalo bone marrow. Bone marrow aspirates were collected from iliac crest of pelvis of buffalo calves immediately after slaughtering of animals. MSCs were separated by density gradient method and cultured in high glucose DMEM supplemented with 15% FBS. Population doubling time and characterization of MSCs was done at 4th passage. Population doubling time for fourth passaged fresh MSCs was 38.63±1.70 hrs. Buffalo MSCs showed positive alkaline phosphatase (AP) activity and positive expression of surface markers (CD73 and CD105) before cryopreservation. MSCs were cryopreserved by slow freezing and fast freezing method for three months. Me2SO and Glycerol were used as cryoprotectants in five different combinations viz. medium I (10% Me2SO), II (7% Me2SO and 3% Glycerol), III (5% Me2SO and 5% Glycerol), IV (3% Me2SO and 7% Glycerol) and V (Commercial serum free medium).for freezing MSCs. Post-cryopreserved MSCs were analyzed for viability, population doubling time and their expression pattern. The results of present study revealed significantly lower population doubling time and higher viability percentage after slow freezing as compared to fast freezing with cryopreservation media II and V. Cryopreserved MSCs maintained the expression pattern of CD73 and CD105 markers similar to fresh MSCs concluding the study that MSCs can be better cryopreserved with cryomedia II and V by slow freezing protocol.
Akram T, Shah RA, Fazili MR, Mudasir H, Mir BA, Hussain SS, Ahmad SM, Dar PA, Ganai NA and Shabir N (2017). Comparative efficiency of goat mesenchymal stem cell isolation from bone marrow and bone chip. Small Ruminant Research 153:87-94.
Bruder SP, Jaiswal N and Haynesworth SE (1997). Growth kinetics, self renewal and osteogenic potential of purified human mesenchymal stem cells during extensive subcultivation and following dryopreservation. Journal of Cell Biochemistry 64(2):278-94.
Campos LL, Landim-Alvarenga FC, Ikeda TL, Monteiro BA, Maia L, Freitas-Dell’Aqua CP and De Vita B (2017). Isolation, culture, characterization and cryopreservation of stem cells derived from amniotic mesenchymal layer and umbilical cord tissue of bovine fetuses. Pesquisa Veterinária Brasileira 36(3):278-286.
Dariolli R, Bassaneze V, Nakamuta JS, Omae SV, Cristina L, Campos G and Krieger JE (2013) Porcine adipose tissue-derived mesenchymal stem cells retain their proliferative characteristics, senescence, karyotype and plasticity after long-term cryopreservation. PLoSONE 8(7):e67939.
Dev K, Giri SK, Kumar A, Yadav A, Singh B and Gautam S (2011). Derivation, characterization and differentiation of buffalo (Bubalus bubalis) amniotic fluid derived stem cells. Reproduction in Domestic Animals 47:704-711.
Doan CC, Truong NH, Vu NB, Nguyen TT, Nguyen HM, Nguyen KG, Do S, Phan NK and Pham PV (2012). Isolation, culture and cryopreservation of human bone marrow derived mesenchymal stem cells. International Journal of Plant, Animal and Environmental Science 2(2):358-367.
Eslaminejad MB, Nazarian H, Falahi F, Taghiyar L and Daneshzadeh MT (2009). Ex vivo expansion and differentiation of mesenchymal stem cells from goat bone marrow. Iranian Journal of Basic Medical Sciences 12(2):70-79.
Gade NE, Pratheesh MD, Nath A, Dubey PK, Amarpal, Sharma B, Saikumar G and Sharma GT (2012). Molecular and cellular characterization of buffalo bone marrow-derived mesenchymal stem cells. Reproduction in Domestic Animals 48(3):10-15.
Hepsibha P, Meenambigai TV, Mangalagowri A, Palanisamy A, Stalin A, Nithya S and Kumanan K (2011). Multipotent differentiation potential of buffalo adipose tissue derived mesenchymal stem cells. Asian Journal of Animal and Veterinary Advances 6:772-788.
Janz FL, Debes AA, Cavaglieri RC, Duarte SA, Romão CM, Morón AF and Zugaib MSP (2012) Evaluation of distinct freezing methods and cryoprotectants for human amniotic fluid stem cells cryopreservation. Journal of Biomedicine and Biotechnology 1155-1165.
Laura C, Groza I, Oana L, Pall E, Tean CP, Catana P and Cenariu M (2008). Canine mesenchymal stem cells isolation from bone marrow aspirates. Bulletin of University of Agricultural Sciences and Veterinary Medicine 65:96-101.
Liu Y, Xu X, Ma XH and Cui ZF (2011). Effect of various freezing solutions on cryopreservation of mesenchymal stem cells from different animal species. Cryo Letters 32(5):425-435.
Lui Y, Xu X, Ma X, Martin RE, Watt S and Cui Z (2010). Cryopreservation of human bone marrow derived mesenchymal stem cells with reduced dimethyl sulphoxide and well-defined freezing solutions. Biotechnology Progress 26:1635-1643.
McCarty RC, Gronthos S, Zannettino AC, Foster BK and Xian CJ (2009). Characterisation and development potential of ovine bone marrow derived mesenchymal stem cells. Journal of Cell Physiology 219:324-33.
Martinello T, Bronzini I, Maccatrozzo L, Iacopetti I, Sampaolesi M, Mascarello F and Patruno M (2010). Cryopreservation does not affect the stem characteristics of multipotent cells isolated from equine peripheral blood. Tissue Engineering Part C 16(4):771-781.
Mitchell A, Rivas KA, Roger SR and Watts AE (2015). Cryopreservation of equine mesenchymal stem cells in 95% autologous serum and 5% DMSO does not alter post-thaw growth or morphology in vitro compared to fetal bovine serum or allogeneic serum at 20 or 95% and DMSO at 10 or 5%. Stem Cell Research and Therapy 6:231-243.
Mehrabani D, Tajedini M, Tamadon A, Dianatpour M, Parvin F, Zare S and Rahmanifar F (2016) Establishment, characterization and cryopreservation of Fars native goat fetal fibroblast cell lines. Asian Pacific Journal of Reproduction 5(3):247-251.
Nair MB, Varma HK, Menon KV, Shenoy SJ and John A (2009). Reconstruction of goat femur segmental defects using triphasic ceramic-coated hydroxyapatite in combination with autologous cells and platelet rich plasma. Acta Biomaterialia 5:1742-1755.
Pavon A, Beloqui I, Salcedo JM and Martin AG (2017). Cryobanking mesenchymal stem cells. Methods in Molecular Biology 1590:191-196.
Peterbauer-Scherb A, van GM, Meinl A, Gabriel C, Redl H and Wolbank S (2010). Isolation of pig bone marrow mesenchymal stem cells suitable for one-step procedures in chondrogenic regeneration. Journal of Tissue Engineering and Regenerative Medicine 4(6):485-490.
Reed SA and Johnson SE (2008). Equine umbilical cord blood contains a population of stem cells that express Oct4 and differentiate into mesodermal and endodermal cell types. Journal of Cell Physiology 215:329-336.
Rentsch C, Hess R, Rentsch B, Hofmann A, Manthey S, Scharnweber D, Biewener A and Zwipp H (2010). Ovine bone marrow mesenchymal stem cells: isolation and characterization of the cells and their osteogenic differentiation potential on embroidered and surface-modiûed polycaprolactone-co-lactide scaffolds. In Vitro Cellular and Developmental Biololgy Animal 46:624-634.
Zhu X, Yuan F, Li H, Zheng Y, Xiao Y and Yan F (2013) Evaluation of canine bone marrow-derived mesenchymal stem cells after long-term cryopreservation. Zoological Science 30 (12):1032-1037.