Murine Mesenchymal Stem Cell Isolation and Characterization

The procedure has been previously described [53 (link)]. Briefly, bone marrow from tibias and femurs of C57BL/6 mice were collected. Cell suspensions were seeded at a concentration of 10⁶ cells/cm² in modified minimum essential Eagle's medium (MEM) supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Hyclone, Thermo Fisher Scientific), 2 mM glutamine, 100 U/ml penicillin, 100 mg/ml streptomycin (Lonza, Levallois-Perret, France) and 2 ng/ml human basic fibroblast growth factor (bFGF) (R&D Systems, Lille, France). MSCs were CD29⁺, Sca1⁺, CD73⁺. Functionally, MSCs had the capacity to differentiate into adipocytes, chondrocytes and osteoblasts when cultured under specific and appropriated conditions [53 (link)].

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Akhter W., Nakhle J., Vaillant L., Garcin G., Le Saout C., Simon M., Crozet C., Djouad F., Jorgensen C., Vignais M.L, & Hernandez J. (2023). Transfer of mesenchymal stem cell mitochondria to CD4+ T cells contributes to repress Th1 differentiation by downregulating T-bet expression. Stem Cell Research & Therapy, 14, 12.

Publication 2023

penicillin streptomycin human basic fibroblast growth factor (bFGF)

Adipocytes Basic fibroblast growth factor Bone marrow C57bl mice Cd73 Cells Chondrocytes Femurs Fetal bovine serum Glutamine Human Osteoblasts Penicillin Sca1 Streptomycin Tibias

Corresponding Organization :

Other organizations : Institute for Regenerative Medicine & Biotherapy, Institut de Génomique Fonctionnelle, Institut de Génétique Moléculaire de Montpellier, Institute for Neurosciences of Montpellier, Hôpital Saint Eloi, Centre Hospitalier Universitaire de Montpellier

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Variable analysis

independent variables

Cell culture conditions (modified minimum essential Eagle's medium (MEM) supplemented with 10% heat-inactivated fetal bovine serum (FBS), 2 mM glutamine, 100 U/ml penicillin, 100 mg/ml streptomycin, and 2 ng/ml human basic fibroblast growth factor (bFGF))

dependent variables

Mesenchymal stem cell (MSC) phenotype (CD29+, Sca1+, CD73+)
Differentiation capacity of MSCs (adipocytes, chondrocytes, osteoblasts)

control variables

Mouse strain (C57BL/6)
Cell seeding density (10^6 cells/cm^2)

controls

Positive control: Differentiation of MSCs under specific and appropriate conditions [53 (link)]
Negative control: Not explicitly mentioned

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