Comparative study on the directed differentiation ability of mouse bone marrow and adipose-derived mesenchymal stem cells

doi:10.11958/20230437

Abstract

Abstract:

Objective To investigate the targeted differentiation ability of mouse bone marrow derived mesenchymal stem cells (BM-MSCs) and adipose-derived mesenchymal stem cells (AD-MSCs). Methods BM-MSCs and AD-MSCs were isolated and cultured from bone marrow of femur and white adipose tissue of groin of C57BL/6J mice respectively, and the two types of cells were induced by osteogenic, chondrogenic and adipogenic differentiation medium respectively. Alizarin red, alcian blue and oil red O staining were used to detect the differentiated degree of osteogenic, chondrogenic and lipogenic differentiation. Real-time fluorescence quantitative PCR (qPCR) was used to identify MSCs and detected expression levels of directed differentiation-related genes Runx2, Sp7 (osteoblast), Sox9, Col2a1 (chondroblast), Pparg and Cebpa (lipogenesis) to determine the directed differentiation ability of cells. Based on gene expression profiles of mouse and human BM-MSCs and AD-MSCs in GEO database GSE43804 and GSE122778, the differentially expressed genes and their enrichment signal pathways were analyzed. Results The cell morphology of BM-MSCs and AD-MSCs obtained by isolation and culture was different, and spindle-shaped morphology was more obvious in AD-MSCs. Both cells expressed CD29, CD44 and CD90, but did not express CD34 and CD45. AD-MSCs showed higher osteogenic and lipogenic differentiation than those of BM-MSCs after directed induction, while chondrogenic differentiation was lower in AD-MSCs than that of BM-MSCs (P<0.05). After directional induction, expression levels of Runx2, Pparg and Cebpa mRNA were higher in AD-MSCs than those in BM-MSCs, and Sox9 mRNA expression levels were lower than those in BM-MSCs (P<0.05). Highly expressed genes of AD-MSCs in mice and human were enriched in PPAR and WNT signaling pathways. Highly expressed genes of BM-MSCs were enriched in cartilage and bone developmental signaling pathways. Conclusion The osteogenic and adipogenic differentiation ability of mouse AD-MSCs is stronger than those of BM-MSCs, while the chondrogenic differentiation ability AD-MSCs is weaker than that of BM-MSCs. The activation status of PPAR, WNT, cartilages and skeletal system development signaling pathways plays an important regulatory role in determining the different directional differentiation potential of AD-MSCs and BM-MSCs.

Key words: mesenchymal stem cells, bone marrow, fats, PPAR gamma, Wnt signaling pathway, osteogenic differentiation, chondrogenic differentiation, adipogenic differentiation

CLC Number:

R329.24

Figures/Tables 13

References 19

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基因名称	引物序列（5′→3′）	产物大小/bp
Gapdh	上游：ATTGTCAGCAATGCATCCTG	102
Gapdh	下游：ATGGACTGTGGTCATGAGCC	102
CD29	上游：CCTGTAACTCCGACGCCTTT	137
CD29	下游：AAGGTCCCCACTCAGCAATG	137
CD34	上游：TTTCCTGATGAACCGTCGCA	165
CD34	下游：GCAGGGTTGTGAGGTACTGT	165
CD44	上游：ACCTTGGCCACCACTCCTAAT	118
CD44	下游：TCACATGGGAGTCTTCACTTGG	118
CD45	上游：CTTTGCTTATGTGGCGTGTGT	139
CD45	下游：TTATCCCCTTCTGATGCGCC	139
CD90	上游：TCCAAGTCGGAACTCTTGGC	133
CD90	下游：TCCAGGCGAAGGTTTTGGTT	133
Runx2	上游：CTCTGCACCAAGTCCTTTTAATC	104
Runx2	下游：AGGAGGGGTAAGACTGGTCATAG	104
Sp7	上游：CTGAGAGAGGAGCAGATCCC	135
Sp7	下游：GTGAGCTTCTTCCTGGGTAGG	135
Sox9	上游：AGGAAGTCGGTGAAGAACGG	167
Sox9	下游：GGACCCTGAGATTGCCCAGA	167
Col2a1	上游：ATGAGGGAGCGGTAGAGACC	190
Col2a1	下游：GCCCTAATTTTCGGGCATCC	190
Pparg	上游：GAGCACTTCACAAGAAATTACC	192
Pparg	下游：GAACTCCATAGTGGAAGCCT	192
Cebpa	上游：CAAGAACAGCAACGAGTACCG	120
Cebpa	下游：AGGCGGTCATTGTCACTGGT	120

基因名称	引物序列（5′→3′）	产物大小/bp
Gapdh	上游：ATTGTCAGCAATGCATCCTG	102
Gapdh	下游：ATGGACTGTGGTCATGAGCC	102
CD29	上游：CCTGTAACTCCGACGCCTTT	137
CD29	下游：AAGGTCCCCACTCAGCAATG	137
CD34	上游：TTTCCTGATGAACCGTCGCA	165
CD34	下游：GCAGGGTTGTGAGGTACTGT	165
CD44	上游：ACCTTGGCCACCACTCCTAAT	118
CD44	下游：TCACATGGGAGTCTTCACTTGG	118
CD45	上游：CTTTGCTTATGTGGCGTGTGT	139
CD45	下游：TTATCCCCTTCTGATGCGCC	139
CD90	上游：TCCAAGTCGGAACTCTTGGC	133
CD90	下游：TCCAGGCGAAGGTTTTGGTT	133
Runx2	上游：CTCTGCACCAAGTCCTTTTAATC	104
Runx2	下游：AGGAGGGGTAAGACTGGTCATAG	104
Sp7	上游：CTGAGAGAGGAGCAGATCCC	135
Sp7	下游：GTGAGCTTCTTCCTGGGTAGG	135
Sox9	上游：AGGAAGTCGGTGAAGAACGG	167
Sox9	下游：GGACCCTGAGATTGCCCAGA	167
Col2a1	上游：ATGAGGGAGCGGTAGAGACC	190
Col2a1	下游：GCCCTAATTTTCGGGCATCC	190
Pparg	上游：GAGCACTTCACAAGAAATTACC	192
Pparg	下游：GAACTCCATAGTGGAAGCCT	192
Cebpa	上游：CAAGAACAGCAACGAGTACCG	120
Cebpa	下游：AGGCGGTCATTGTCACTGGT	120

细胞	CD29/×10^-3	CD44/×10^-3	CD90/×10^-3	CD34	CD45
BM-MSCs	1.861±0.288	0.562±0.120	0.456±0.017	0	0
AD-MSCs	0.805±0.208	0.433±0.158	0.851±0.261	0	0

细胞	CD29/×10^-3	CD44/×10^-3	CD90/×10^-3	CD34	CD45
BM-MSCs	1.861±0.288	0.562±0.120	0.456±0.017	0	0
AD-MSCs	0.805±0.208	0.433±0.158	0.851±0.261	0	0

细胞	诱导分化前		诱导分化后
细胞	Runx2/×10^-5	Sp7/×10^-6	Runx2/×10^-4	Sp7/×10^-5
BM-MSCs	4.85±0.99	6.11±1.17	2.42±0.44	4.66±3.73
AD-MSCs	4.36±1.13	2.01±1.22	4.61±0.92	4.17±3.54
t	0.510	3.024^＊	3.923^＊	1.171