• SANKARI D SRM University


Objective: Goal of this study is to find a simple method for isolation, colony formation and Trilineage differentiation of Mesenchymal stem cells from
bone marrow of Wistar Rat. Adherent capacity, morphology, trilineage differentiation and colony formation of bone marrow mesenchymal stem cells
were studied in low glucose, high glucose Dulbecco modified eagle medium with various concentration of fetal bovine serum.
Methods: Mesenchymal stem cells were isolated from bone marrow of Wistar rat by Ficoll (sigma) density gradient centrifugation with plastic
adherence method. Bone marrow was collected from femur and Tibia of 6-weeks-old Wistar rat; Bone marrow was cultured in Dulbecco's modified
Eagles's medium (DMEM) with low glucose, high glucose supplement (Invitrogen) and 10%, 15% of fetal bovine serum (FBS) at the density of
1 × 10
, incubated at 37°C in 5% of CO
; adherent capacity, colony formation were studied. Differentiation of bone marrow mesenchymal stem cells
(BMMSCs) was monitored under suitable differentiation medium. The cells were refeed every 3-4 days and passaged when the cells reached 80-90%
Result: BMMSCs were adhered in tissue culture flask at 24 hrs, 48 hrs in high glucose and low glucose DMEM with 10% of FBS, respectively. Colony
formation was faster in high glucose and low glucose DMEM with 15% of FBS compared to high glucose and low glucose DMEM with10% FBS.
Morphological changes were observed in BMMSCs high glucose, low glucose DMEM with 10% FBS, but no changes were found in the differentiation
of BMMSCs in high glucose, low glucose DMEM. Differentiation of BMMSCs was nourishing in third passage cells.
Conclusion: High glucose DMEM with 10% FBS is a good supplement for adherence of cells whereas high glucose, low glucose DMEM with 15% FBS
can be utilized for rapid Colony formation. Third passage BMMSCs is fruitful for differentiation of BMMSCs.
Keywords: Wistar rat, Bone marrow, Mesenchymal stem cells, Adherent capacity, Colony formation, Differentiation, Osteoblast, Chondrocytes,

Author Biography

SANKARI D, SRM University

Head, Department of Biotech


Friedenstein AJ, Gorskaja JF, Kulagina NN. Fibroblast precursors in

normal and irradiated mouse hematopoietic organs. Exp Hematol


Beyer Nardi N, da Silva Meirelles L. Mesenchymal stem cells: Isolation,

in vitro expansion and characterization. Handb Exp Pharmacol


Prockop DJ. Marrow stromal cells as stem cells for nonhematopoietic

tissues. Science 1997;276(5309):71-4.

Pittenger MF, Marshak DR. In: Marshak DR, Gardner RL, Gottlieb D,

editors. Stem Cell Biology. New York: Cold Spring Harbor Laboratory

Press, Cold Spring Harbor; 2001.

Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F,

Krause D, et al. Minimal criteria for defining multipotent mesenchymal

stromal cells. The international society for cellular therapy position

statement. Cytotherapy 2006;8(4):315-7.

Kang XQ, Zang WJ, Song TS, Xu XL, Yu XJ, Li DL, et al. Rat bone

marrow mesenchymal stem cells differentiate into hepatocytes in vitro.

World J Gastroenterol 2005;11(22):3479-84.

Oswald J, Boxberger S, Jørgensen B, Feldmann S, Ehninger G,

Bornhäuser M, et al. Mesenchymal stem cells can be differentiated into

endothelial cells in vitro. Stem Cells 2004;22(3):377-84.

Polisetti N, Chaitanya VG, Babu PP, Vemuganti GK. Isolation,

characterization and differentiation potential of rat bone marrow

stromal cells. Neurol India 2010;58(2):201-8.

Ayatollahi M, Salmani MK, Geramizadeh B, Tabei SZ, Soleimani M,

Sanati MH. Conditions to improve expansion of human mesenchymal

stem cells based on rat samples. World J Stem Cells 2012;4(1):1-8.

Meirelles Lda S, Nardi NB. Murine marrow-derived mesenchymal stem

cell: Isolation, in vitro expansion, and characterization. Br J Haematol


Yameen Z, Leavesley Z, Upton Z, Xiao Y. Multileneage differentiation

potential of bone and cartilage cells derived from explant culture. Open

Stem Cell J 2009;1:10-9.

Wilson A, Trumpp A. Bone-marrow haematopoietic-stem-cell niches.

Nat Rev Immunol 2006;6(2):93-106.

Fu WL, Zhang JY, Fu X, Duan XN, Leung KK, Jia ZQ, et al.

Comparative study of the biological charecteristics of mesenchymal

stem cells from bone marrow and peripheral blood of rats. Tissue Eng

Part A 2012;18(17-18):1793-803.

Taran R, Mamidi MK, Singh G, Dutta S, Parhar IS, John JP, et al.

In vitro and in vivo neurogenic potential of mesenchymal stem cells

isolated from different sources. J Biosci 2014;39(1):157-69.



How to Cite

D, S., T. P, A. D, and T. KANNAN P. “A NOVEL METHOD FOR ISOLATION AND TRILINEAGE DIFFERENTIATION OF RAT BONE MARROW DERIVED MESENCHYMAL STEM CELLS”. Asian Journal of Pharmaceutical and Clinical Research, vol. 8, no. 5, Sept. 2015, pp. 126-9, https://innovareacademics.in/journals/index.php/ajpcr/article/view/6933.



Original Article(s)