Fudan University Journal of Medical Sciences

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A method for preparing efficient adipogenic differentiation of mice adipose stem cells

Qiu-kai TANG, Sai-sai SONG, Yan TANG

Fudan University Journal of Medical Sciences, 2025, 52(02): 277-284, 291. DOI: 10.3969/j.issn.1672-8467.2025.02.015

cDNA	F primer（5’-3’）	R pimer（5’-3’）
PDGFRα	GAGGAAGCCGAGATACCCC	TGCTGTGGATCTGACTTCGAG
PDGFRβ	AGGACAACCGTACCTTGGGTGACT	CAGTTCTGACACGTACCGGGTCTC
DPP4	ACCGTGGAAGGTTCTTCTGG	CACAAAGAGTAGGACTTGACCC
CD142	AACCCACCAACTATACCTACACT	GTCTGTGAGGTCGCACTCG
ICAM1	TGCCTCTGAAGCTCGGATATAC	TCTGTCGAACTCCTCAGTCAC
Pref1	AGTGCGAAACCTGGGTGTC	GCCTCCTTGTTGAAAGTGGTCA
18s	CGCCGCTAGAGGTGAAATTCT	CATTCTTGGCAAATGCTTTCG
CD31	ACGCTGGTGCTCTATGCAAG	TCAGTTGCTGCCCATTCATCA
SM22	CAACAAGGGTCCATCCTACGG	ATCTGGGCGGCCTACATCA

Tab 1 Primers for qPCR used in this study

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Fig 1 Characteristics of adipose-derived stem cells from subcutaneous fat of mice treated with different digestion times A: Illustrates the condition of adipose tissue obtained at different digestion durations (top), the morphology of adherent adipose-derived stem cells (ADSCs) cultured under different digestion times (middle), bodipy staining showing adipogenesis under different digestion times (bottom); B: Quantification of cell number under different digestion time; C: Quantification of adipogenic differentiation from A. Data were expressed as $\bar x \pm s$, ⁽¹⁾P < 0.05, ⁽²⁾P < 0.01, ⁽³⁾P < 0.001;ns: Not significant.
Fig 2 Adipogenic differentiation of adipose-derived stem cells from subcutaneous fat of mice treated with different digestion times A: Microscopic imaging after oil red O staining of adipocyte with different induction conditions; B: mRNA expression levels of mature adipocyte-related genes PPARγ and Ap2; C: Expression levels of mature adipocyte-related proteins PPARγ and Ap2 via Western blot analysis. Data were expressed as $\bar x \pm s$, ⁽¹⁾ P < 0.01, ⁽²⁾ P < 0.001.
Fig 3 Vascular stromal component in mouse subcutaneous adipose tissue A: Quantification of CD34+ population abundance in SVF from mice; B: Quantification of population abundance in CD31-CD45-cells; C: Schematic representation of adipose tissue composition, including mature adipocytes, mesenchymal cells and vascular cells; D: Expression levels of vascular-related proteins CD31 and SM22 in mesenchymal cells and vascular cells; E: Detection of expression levels of vascular-related protein CD31 in mesenchymal cells and vascular cells via Western blot; F: Similar expression levels of preadipocyte-related genes PDGFR and Pref1 in mesenchymal cells and vascular cells; G: Differential expression of preadipocyte-related genes DPP4 and PPARγ in mesenchymal cells and vascular cells. Data were expressed as $\bar x \pm s$, ⁽¹⁾P < 0.05, ⁽²⁾ P < 0.01, ⁽³⁾ P < 0.000 1;ns: Not significant.
Fig 4 Adipogenic differentiation of stromal cells and vascular-related cells isolated from mouse subcutaneous adipose tissue in vitro A: Microscopic images post Oil Red O staining of mesenchymal cell and vascular cell; B: Expression levels of mature adipocyte-related genes PPARγ and Ap2; C: Expression levels of mature adipocyte-related protein PPARγ and Ap2 via Western blot analysis. ⁽¹⁾ P < 0.000 1.

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