Maternal inappropriate calcium intake aggravates dietary-induced obesity in male offspring by affecting the differentiation potential of mesenchymal stem cells

Table 1 Details of diet formulations in this study (g/kg diet)

Ingredients (g)	Reproductive diets				Feeding diets (0.70%)
	Deficient calcium diet (0.05%)	Low calcium diet (0.25%)	Normal calcium diet (0.70%)	High calcium diet (1.20%)	Normal fat diet (H10010)	High fat diet (H10060)
Casein	200.00	200.00	200.00	200.00	189.58	258.45
Cystine	3.00	3.00	3.00	3.00	2.84	3.88
Cornstarch	396.30	391.30	380.00	367.50	298.59	161.53
Maltodextrin	132.00	132.00	132.00	132.00	33.18	88.91
Sucrose	100.00	100.00	100.00	100.00	331.77	---
Fibrin	50.00	50.00	50.00	50.00	47.40	64.61
Soybean oil	70.00	70.00	70.00	70.00	23.70	32.31
Lard oil	---	---	---	---	18.96	316.60
Mineral mixture without calcium (M1004)	35.00	35.00	35.00	35.00	---	---
Mineral mixture (M1002)	---	---	---	---	9.48	12.92
Calcium bicarbonate	---	---	---	---	12.32	16.80
Calcium carbonate (CaCO3)	1.25	6.25	17.50	30.00	5.21	7.11
Potassium citrate. H2O	---	---	---	---	15.64	21.32
Vitamin mixture (V1002)	10.00	10.00	10.00	10.00	9.48	12.92
Choline Bitartrate	2.50	2.50	2.50	2.50	1.90	2.58
Antioxidant (TBHQ)	0.014	0.014	0.014	0.014	0.047	0.065

Table 2 Effects of maternal different calcium intake on cell cycle and purity of bone marrow mesenchymal stem cells among male offspring

Indicators (%)	NC group (n = 9)	DC group (n = 9)	LC group (n = 9)	HC group (n = 9)	χ2	P value
Cell cycle
G0G1 Phase	91.27 ± 3.14	94.26 ± 2.09	95.08 ± 2.54	92.46 ± 5.01	1.774	0.939
G2M Phase	0.85 ± 0.0091	0.94 ± 0.011	1.05 ± 0.013	0.79 ± 0.0082
S Phase	7.89 ± 0.81	4.81 ± 0.56	3.87 ± 0.75	6.75 ± 0.94
Flow cytometry
Sca-1 (+)	95.03 ± 4.85	98.72 ± 6.12	99.31 ± 5.48	95.49 ± 3.84	4.788	0.188
CD90 (+)	98.12 ± 7.15	99.15 ± 2.04	99.07 ± 8.12	99.31 ± 6.74	0.608	0.895
CD29 (+)	99.13 ± 6.48	99.90 ± 3.46	97.02 ± 8.07	99.62 ± 7.01	0.549	0.908
CD34 (-)	99.08 ± 7.42	96.67 ± 2.93	97.94 ± 1.48	97.29 ± 1.92	0.364	0.546
CD31 (-)	99.46 ± 6.82	99.42 ± 4.12	98.94 ± 1.38	99.26 ± 1.57	0.159	0.690
CD45 (-)	99.79 ± 1.08	99.83 ± 2.54	98.48 ± 1.12	98.65 ± 1.75	0.287	0.963
CD49d (-)	99.13 ± 6.21	98.16 ± 2.32	99.06 ± 2.98	98.11 ± 1.09	5.087	0.166

Four-week-old C57BL/6J female mice were respectively fed with deficient (DC, 0.05%), low (LC, 0.25%), normal (NC, 0.70%) and high (HC,1.20%) calcium reproductive diets for 6 wk before mating and continued their diets throughout gestation and lactation. After weaning, male offspring at 7-day-old from DC, LC, NC and HC groups were chosen to determine the cell cycle and purity of bone marrow mesenchymal stem cells (1 × 10⁶) by flow cytometry. All values were shown as mean ± standard deviation (n = 9/group), and comparisons were made by χ² test. NC: Normal-calcium reproductive diet; DC: Deficient-calcium reproductive diet; LC: Low-calcium reproductive diet; HC: High-calcium reproductive diet.