Comparative study of biological characteristics of mesenchymal stem cells isolated from mouse bone marrow and peripheral blood

Abstract

Mesenchymal stromal cells (MSCs) possess self-renewal and multilineage differentiation potential, indicating their prospects as cellular therapeutic agents for regenerative medicine. Although adult bone marrow (BM) is the major source of these cells for clinical use, harvesting requires invasive procedures. Therefore, alternative sources, such as peripheral blood (PB), are needed. The objective of the current study was to compare PB-MSCs and BM-MSCs with regard to their biological characteristics. PB-MSCs and BM-MSCs were isolated from 4-week-old BALB/c white mice by density gradient centrifugation and cultured in DMEM + 10% fetal bovine serum until passage four. Morphological features, proliferation, cell surface marker expression and trilineage differentiation potential were assessed for both PB-MSCs and BM-MSCs. No significant differences in morphological features were observed. BM-MSCs had a higher proliferative capability than PB-MSCs as measured by XTT assays. Both PB-MSCs and BM-MSCs had broadly similar cell surface marker expression, but PB-MSCs had positive expression of cluster of differentiation (CD)146 and CD140b. Both PB-MSCs and BM-MSCs were capable of trilineage differentiation. Although BM-MSCs had a greater capacity for osteogenic and chondrogenic differentiation than PB-MSCs, PB-MSCs had a better capability for adipogenic differentiation than BM-MSCs. In conclusion, PB-MSCs and BM-MSCs have very similar biological characteristics. Thus, PB is a promising source for easily obtaining MSCs in mice.

Keywords: BALB/c mice; bone marrow; bone marrow-mesenchymal stromal cells; mesenchymal stem cells; peripheral blood; peripheral blood-mesenchymal stromal cells.

Figure 1.

XTT assays of BM-MSCs and PB-MSCs to measure the proliferation rate. Both MSCs were seeded at 125, 250, 500 and 1,000 cells/well and allowed to grow for 7 days then the optical density was measured. The proliferation rate of BM-MSCs was higher than that of PB-MSCs and it was directly proportional to the number of inoculated cells. ^*P<0.05 vs. PB-MSCs. BM-MSCs, bone marrow-mesenchymal stromal cells; PB, peripheral blood; OD, optical density.

Figure 2.

Flow cytometric analysis of BM-MSCs and PB-MSCs. BM-MSCs were positive CD29 and negative for other markers, while PB-MSCs were positive for CD146, CD29, and CD140b and negative for Sca-1, CD44, CD45, CD90 and CD105. CD, cluster of differentiation; BM-MSCs, bone marrow-mesenchymal stromal cells; PB, peripheral blood; FITC, fluorescein isothiocyanate; PE, phycoerythrin.

Figure 3.

Differentiation capability of BM-MSCs and PB-MSCs. Osteogenic differentiation of (A) BM-MSCs and (B) PB-MSCs. Both stained with Alizarin Red (magnification, x10). Adipogenic differentiation of (C) BM-MSCs and (D) PB-MSCs stained with Oil red O (magnification, x10). Chondrogenic differentiation of (E) BM-MSCs and (F) PB-MSCs stained with toluidine blue (magnification, x4). BM-MSCs, bone marrow-mesenchymal stem cells; PB, peripheral blood.

Figure 4.

Quantitative analysis of the differentiation capability of BM-MSCs and PB-MSCs. (A) Osteogenesis detection by Ca-ion assays. (B) The adipogenesis percentage in PB-MSCs and BM-MSCs. (C) Chondro detection by GAG assay. Data are shown as the mean ± standard deviation (n=3). ^*P<0.05 vs. the control, ^†P<0.05 vs. BM-MSCs, ^‡P<0.05 vs. PB-MSCs. BM-MSCs, bone marrow-mesenchymal stromal cells; PB, peripheral blood; chondro, chondrogenesis.