Umbilical cord-derived mesenchymal stem cells cultured in the MCL medium for aplastic anemia therapy

Abstract

Background: Mesenchymal stem cells (MSCs) are a class of adult stem cells with self-renewal and multidirectional differentiation potential that may be a treatment for aplastic anemia (AA).

Method: Umbilical cord-derived MSCs were cultured in three media (Mesencult-XF, MCL, and StemPro MSC SFM CTS). HGF, PGE2, ANG-1, TGF-β1, IFN-γ, and TNF-α were detected using ELISA. The AA mouse model was built via post-irradiation lymphocyte infusion. After different treatments, routine blood, VEGF, and Tregs were detected every week. On day 28, all mice were killed, and their femurs were stained with HE.

Results: Umbilical cord-derived MSCs cultured in the three media all conformed to the general characteristics of MSCs. HGF secreted by MSCs in the Mesencult-XF, and MCL was greater than that in the StemPro MSC SFM CTS; ANG-1 and TGF-β1 in the MCL were more than that in Mesencult-XF and StemPro MSC SFM CTS; PGE2 in the MCL and StemPro MSC SFM CTS was more than that in the Mesencult-XF. MSCs in the MCL and StemPro MSC SFM CTS inhibited IFN-γ and TNF-α more than those in the Mesencult-XF. The peripheral blood cell in the AA groups was at a low level while that in the MSC group recovered rapidly. The Treg ratio and VEGF level in the MSC group were higher than those in the AA group. The bone marrow (BM) recovered significantly after MSC infusion.

Conclusion: MSCs in the MCL were advantageous in supporting hematopoiesis and modulating immunity and had the potential for effective treatment of AA.

Keywords: Aplastic anemia; Mesenchymal stem cell; Umbilical cord; Wharton's jelly.

Fig. 1

Morphology and differentiation of MSCs. A Morphology of MSCs in different media, scale bar: 25 μm; B Differentiation of MSCs in different media, scale bar: 50 μm (1. Mesencult-XF, 2. MCL, 3. StemPro MSC SFM CTS)

Fig. 2

Immunophenotype of MSCs in different media (1. Mesencult-XF; 2. MCL; 3. StemPro MSC SFM CTS)

Fig. 3

Cytokines (HGF, ANG-1, PGE2, and TGF-β1) secreted by MSCs in different media (**P < 0.01)

Fig. 4

Establishment of AA mouse model and treatment of MSCs in cultured in the MCL medium. A Schematic illustration of the animal experiments; B Survival curve of mice in the three groups; C VEGF levels in the three groups of mice at different time points (*P < 0.05; **P < 0.01)

Fig. 5

WBC, Hb and PLT counts of mice in the three groups at different time points A WBC count; B Hb count; C PLT count (*P < 0.05)

Fig. 6

Tregs of mice in the irradiation, AA, MSC groups at different time points. A Flow cytometry analysis of Treg change in the irradiation group; B Flow cytometry analysis of Treg change in the AA group; C Flow cytometry analysis of Treg change in the MSC group; D Quantitative comparison of Treg differences among the three groups (*P < 0.05)

Fig. 7

HE staining of the femur of mice in the irradiation, AA, MSC groups at different magnifications (Scale bar at left: 50 μm; Scale bar at right: 100 μm)