ZOOLOGICAL RESEARCH
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2023
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IF 4.9
Cong Li,Rui Bi,Lin Wang,Yu-Hua Ma,Yong-Gang Yao,Ping Zheng
Tree shrews ( Tupaia belangeri chinensis ) share a close relationship to primates and have been widely used in biomedical research. We previously established a spermatogonial stem cell (SSC)-based gene editing platform to generate transgenic tree shrews. However, the influences of long-term expansion on tree shrew SSC spermatogenesis potential remain unclear. Here, we examined the in vivo spermatogenesis potential of tree shrew SSCs cultured across different passages. We found that SSCs lost spermatogenesis ability after long-term expansion (>50 passages), as indicated by the failure to colonize the seminiferous epithelium and generate donor spermatogonia (SPG)-derived spermatocytes or spermatids marking spermatogenesis. RNA sequencing (RNA-seq) analysis of undifferentiated SPGs across different passages revealed significant gene expression changes after sub-culturing primary SPG lines for more than 40 passages on feeder layers. Specifically, DNA damage response and repair genes (e.g., MRE11 , SMC3 , BLM , and GEN1 ) were down-regulated, whereas genes associated with mitochondrial function (e.g., NDUFA9 , NDUFA8 , NDUFA13 , and NDUFB8 ) were up-regulated after expansion. The DNA damage accumulation and mitochondrial dysfunction were experimentally validated in high-passage cells. Supplementation with nicotinamide adenine dinucleotide (NAD + ) precursor nicotinamide riboside (NR) exhibited beneficial effects by reducing DNA damage accumulation and mitochondrial dysfunction in SPG elicited by long-term culture. Our research presents a comprehensive analysis of the genetic and physiological attributes critical for the sustained expansion of undifferentiated SSCs in tree shrews and proposes an effective strategy for extended in vitro maintenance.
研究领域:
生物医学研究
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干细胞生物学
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分子遗传学
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生殖生物学
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DOI
PubMed
高影响因子
IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL
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2023
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IF 2.1
Ke Shenghui,Feng Yiwei,Luo Liying,Qin Wanzhao,Liu Huayu,Nie Jingchong,Liang Beijiang,Ma Hongjie,Xie Mao,Li Jingyu,Niu Zhijie,Li Guojian,Tang Anzhou,Xia Wei,He Guangyao
A method for the in vitro isolation, purification, identification, and induced differentiation of satellite cells from adult tree shrew skeletal muscle was established. The mixed enzyme digestion method and differential adhesion method were used to obtain skeletal muscle satellite cells, which were identified and induced to differentiate to verify their pluripotency. The use of a mixture of collagenase II, hyaluronidase IV, and DNase I is an efficient method for isolating adult tree shrew skeletal muscle satellite cells. The P3 generation of cells had good morphology, rapid proliferation, high viability, and an “S”-shaped growth curve. Reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence staining indicated that marker genes or proteins were expressed in skeletal muscle satellite cells. After myogenic differentiation was induced, multiple-nucleated myotubes were observed, and the MyHC protein was expressed. The expression of myogenic marker genes changed with the differentiation process. After the induction of adipogenic differentiation, orange-red lipid droplets were observed, and the expression of adipogenic marker genes increased gradually with the differentiation process. In summary, satellite cells from adult tree shrew skeletal muscle were successfully isolated using a mixed enzyme digestion method, and their potential for differentiation into myogenic and adipogenic cells was confirmed, laying a foundation for further in vitro study of tree shrew muscle damage.
研究领域:
细胞生物学
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肌肉生理学
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干细胞研究
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产品关联
DOI
PubMed
高影响因子
LIFE SCIENCES
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2023
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IF 6.1
Jiahong Feng,Ting Jia,Yue Ren,Hao Zhang,Wanlong Zhu
Aims Leptin is a signaling protein secreted by white adipose tissue encoded by the obesity gene, and its main function is to regulate the food intake and energy metabolism in mammals. Previous studies had found that animal leptin concentration was positively correlated with its body fat, but the leptin concentration of Tupaia belangeri was negatively correlated with its body fat mass . The present study attempted to investigate the mechanisms of leptin concentration negatively correlated with its body fat mass in T. belangeri. Material and methods We measured the leptin concentration of the two groups of animals by enzyme linked immunosorbent assay (ELISA) and quantified the leptin mRNA expression by qPCR. Then, the histological, transcriptomic , and bisulfite sequencing of the two groups of animals were studied. Moreover, to investigate the energy metabolism under the negative correlation, we also analyzed the metabolomics and metabolic rate in T. belangeri . Key findings We revealed the negative correlation was mediated by leptin gene methylation of subcutaneous adipose tissue . Further, we also found that T. belangeri increased energy metabolism with leptin decreased. Significance We challenge the traditional view that leptin concentration was positively correlated with body fat mass, and further revealed its molecular mechanism and energy metabolism strategy. This special leptin secretion mechanism and energy metabolism strategy enriched our understanding of energy metabolism of animals, which provided an opportunity for the clinical transformation of metabolic diseases .
研究领域:
内分泌学
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分子生物学
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代谢学
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动物生理学
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DOI
高影响因子
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