Effects of metformin on cell morphology, proliferation, surface markers and cell cycle of human umbilical cord mesenchymal stem cells

doi:10.3877/cma.j.issn.2095-1221.2019.04.006

Abstract

Abstract:

Objective

To study the effects of different concentrations of metformin on morphology, proliferation, surface markers and cell cycle of human umbilical cord mesenchymal stem cells (hUC-MSC) .

Methods

hUC-MSC were isolated from the umbilical cord of full-term newborns in vitro for subculture. The 6th generation of hUC-MSCs in logarithmic growth phase were used in the study. The control group was compared with different concentrations of metformin (0.1, 1, 5, 10, 20 mmol/L) to observe the morphology of cells treated with different concentrations of metformin. The effects of proliferation rate (MTT method) , cell surface markers and cell cycle (flow cytometry analysis) were analyzed by One-Way ANOVA and LSD-t test.

Results

(1) METF was 0.1?mmol/L, 1?mmol/L, and the cell morphology did not change significantly. When the drug concentration was 5?~ 20?mmol/L, the cell morphology changed more obviously with the increase of drug concentration and culture time. (2) When METF was 0.1?mmol/L (highest 104.06±1.76) , it promoted the proliferation of hUC-MSC. The drug concentration of 1?~ 10?mmol/L could increase the proliferation rate of mesenchymal stem cells in the early stage of culture. With the prolongation of culture time, the cells proliferation was gradually suppressed. METF was 20?mmol/L (24?h: 86.64±0.66, 48?h: 58.38±2.52, 72?h: 17.75±1.35) to inhibit cell proliferation, and the inhibition increased with time (P?< 0.05) . When the concentration of metformin was 5, 10, 20?mmol/L, the expression of CD105 gradually decreased with the increase of drug concentration (F?= 17.539, P?< 0.05) . Metformin did not affect CD44 and CD90. When metformin was 0.1?mmol/L, the ratio of G0/?G1 phase was decreased (64.16±1.20, P?< 0.05) , which promoted the proliferation of mesenchymal stem cells. With the increase of drug concentration, cell proliferation was gradually inhibited.

Conclusion

Metformin can promote the proliferation of hUC-MSC at the concentration of 0.1?mmol/L, and inhibit the proliferation of hUC-MSC and the expression of CD105 at the concentration of 5?~ 20?mmol/L. All different concentrations of metformin would influence the expressions of CD44 and CD90.

Key words: Umbilical cord mesenchymal stem cells, Metformin, Biological therapy, Cell proliferation

Xun Gong, Sheng Yun. Effects of metformin on cell morphology, proliferation, surface markers and cell cycle of human umbilical cord mesenchymal stem cells[J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2019, 09(04): 224-229.

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References 23

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分组	24 h	48 h	72 h
对照组	100	100	100
0.1 mmol/L组	101.28±0.98	104.06±1.76^a	101.51±0.67
1 mmol/L组	116.03±1.82^b	84.20±1.60^b	70.98±2.47^b
5 mmol/L组	115.13±3.06^b	64.64±1.20^b	43.83±2.38^b
10 mmol/L组	108.37±1.26^b	59.69±1.90^b	25.58±0.61^b
20 mmol/L组	86.64±0.66^b	58.38±2.52^b	17.75±1.35^b
F值	140.275	442.340	1668.204
P值	< 0.01	< 0.01	< 0.01

分组	24 h	48 h	72 h
对照组	100	100	100
0.1 mmol/L组	101.28±0.98	104.06±1.76^a	101.51±0.67
1 mmol/L组	116.03±1.82^b	84.20±1.60^b	70.98±2.47^b
5 mmol/L组	115.13±3.06^b	64.64±1.20^b	43.83±2.38^b
10 mmol/L组	108.37±1.26^b	59.69±1.90^b	25.58±0.61^b
20 mmol/L组	86.64±0.66^b	58.38±2.52^b	17.75±1.35^b
F值	140.275	442.340	1668.204
P值	< 0.01	< 0.01	< 0.01

分组	CD44	CD90	CD105
对照组	0.75±0.18	96.96±0.44	72.80±2.90
0.1 mmol/L组	0.53±0.25	96.93±0.89	76.75±2.15
1 mmol/L组	0.53±0.08	97.54±0.22	72.88±2.37
5 mmol/L组	0.50±0.24	97.21±0.47	66.52±2.70^a
10 mmol/L组	0.60±0.14	97.47±0.31	66.74±4.33^a
20 mmol/L组	0.62±0.28	97.67±0.22	57.25±6.92^b
F值	0.731	1.232	17.539
P值	> 0.05	> 0.05	< 0.01

分组	CD44	CD90	CD105
对照组	0.75±0.18	96.96±0.44	72.80±2.90
0.1 mmol/L组	0.53±0.25	96.93±0.89	76.75±2.15
1 mmol/L组	0.53±0.08	97.54±0.22	72.88±2.37
5 mmol/L组	0.50±0.24	97.21±0.47	66.52±2.70^a
10 mmol/L组	0.60±0.14	97.47±0.31	66.74±4.33^a
20 mmol/L组	0.62±0.28	97.67±0.22	57.25±6.92^b
F值	0.731	1.232	17.539
P值	> 0.05	> 0.05	< 0.01

分组	G0/G1（﹪）	S（﹪）	G2/M（﹪）
对照组	66.40±2.65	29.03±3.19	3.56±2.20
0.1 mmol/L组	64.16±1.20^b	29.47±1.00	6.37±0.40^a
1 mmol/L组	72.89±1.14^b	24.19±2.16^a	2.34±1.02
5 mmol/L组	73.59±1.58^b	20.92±0.77^b	5.48±0.84
10 mmol/L组	77.55±1.25^b	16.32±1.10^b	6.18±1.01^a
20 mmol/L组	83.85±1.68^b	13.68±1.67^b	2.47±0.23
F值	122.456	1.232	17.539
P值	< 0.01	< 0.01	> 0.05

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