大鼠白色和棕色脂肪来源的间充质干细胞成脂分化特性的比较

doi:10.16098/j.issn.0529-1356.2016.05.005

摘要/Abstract

摘要：

目的探讨白色脂肪组织（WAT）和棕色脂肪组织（BAT）来源的间充质干细胞成脂分化特性的差异。方法雄性SD大鼠15只，3只用于细胞分离培养，12只用于细胞移植。体外分离培养WAT和BAT两种来源的脂肪干细胞，用油红O染色检测两种干细胞的成脂分化率，用免疫荧光技术检测成脂诱导、分化后的细胞是棕色脂肪细胞还是白色脂肪细胞。4’6-二脒基-2-苯基吲哚（DAPI）标记两种来源的干细胞后移植至腹股沟区，分别在第1、2、3周取材，免疫荧光技术检测植入细胞的分化趋向。结果 WAT来源的干细胞增殖速度明显快于BAT来源的干细胞，前者成脂分化率为0.205±0.069，后者为0.165±0.053，两种干细胞的成脂诱导率差异无统计学意义（P>0.05）。两种干细胞分别植入体内后，在第1、2、3周发现，两种干细胞均表达棕色脂肪特异性蛋白解耦联蛋白1（UCP1）。结论 WAT和BAT来源的干细胞在体外及体内诱导成脂后均分化为棕色脂肪细胞。

关键词: 解耦联蛋白1, 棕色脂肪组织, 白色脂肪组织, 脂肪间充质干细胞, 成脂诱导, 免疫荧光, 大鼠

Abstract:

Objective To study differences of adipogenic differentiation characteristics between the mesenchymal stem cells from white adipose tissue (WAT) and brown adipose tissue (BAT). Methods Fifteen male SD rats were used in this study, 3 for cell separation and 12 for cell transplantation. Adipose mesenchymal stem cells from WAT and BAT were separated and cultured in vitro. Oil red O staining was used to test the adipogenic differentiation rate of each kind of stem cells. Immunofluorescence techniques was used to test the adipogenic differentiation rate of each kind of stem cells, and to detect whether the cells were white fat cells or brown fat cells after adipogenic induction and differentiation. Both stem cells after 4’6-diamidino-2-phenylindole(DAPI) staining were transplanted to the groin area. Sample tissues were obtained in 1, 2 and 3 weeks. Immunofluorescence technique was used to detect the differentiation trend of the implanted cells. Results Proliferation rate of stem cells from WAT was significantly faster than that of BAT. Adipogenic differentiation rate of the former was 0.205±0.069, while the latter was 0.165±0.053. Adipogenic rates of the two kinds of stem cells had no statistical significance (P>0.05). After adipogenic cells were implanted in the body, brown adipose specific proteins uncoupling protein 1(UCP1) were expressed from two kinds of stem cells in 1, 2 and 3 weeks’ tissues. Conclusion Both stem cells of WAT and BAT cultured in vitro and in vivo were differentiated into brown fat cells after adipogentic induction.

Key words: Uncoupling protein 1, Brown adipose tissue, White adipose tissue, Adipose-derived mesenchymal stem cell, Adipogenic induction, Immunofluorescence, Rat

牛艳菲赵国安常玉巧甘黎殷国田郭志坤. 大鼠白色和棕色脂肪来源的间充质干细胞成脂分化特性的比较[J]. 解剖学报, 2016, 47(5): 607-613.

NIU Yan-fei ZHAO Guo-an CHANG Yu-qiao GAN Li YIN Guo-tian GUO Zhi-kun. Differences of adipogentic differention characteristics between the mesenchymal stem cells from white adipose tissue and brown adipose tissue in rats[J]. Acta Anatomica Sinica, 2016, 47(5): 607-613.

参考文献

［1］Jang H,Cho K,Park H,et al. Adipose tissue-derived stem cells for cell therapy of airway allergic diseases in mouse ［J］.Acta Histochem,2011,113(5):501-507.

［2］Alt Eu,Senst C,Murthy SN, et al. Aging alters tissue resident mesenchymal stem cell properties ［J］. Stem Cell Res,2012,8(2):215-225.

［3］Vishnubalaji R, Al-Nbaheen M, Kadalmani B,et al. Comparative investigation of the differentiation capability of bone-marrow-and adipose-derived mesenchymal stem cells by qualitative and qualitative analysis ［J］. Cell Tissue Res,2012,347(2):419-427.

［4］Seale P, Bjork B, Yang W,et al. PRDM 16 controls a brown fat/skeletal muscle switch ［J］. Nature, 2008,454(7207):961-967.

［5］Oliver P, Sanchez J, Caimari A,et al. The intake of a high-fat diet triggers higher brown adipose tissue UCP1 levels in male rats bu not in females［J］. Genes Nutr,2007,2 (1):125-126.

［6］Harms M, Seale P. Brown and beige fat: development, functions and therapeutic potential［J］. Nat Med,2013(19):1252-1263.

［7］Labbé SM, Caron A, Bakan I,et al. In vivo measurement of energy substrate contribution to cold-induced brown adipose tissue thermogenesis［J］. FASEB J, 2015,29(5):2046-2058.

［8］Cypess AM, Weiner LS, Roberts-Toler C,et al. Activation of human brown adipose tissue by a β3-adrenergic receptor agonist［J］. Cell Metab,2015,21(1):33-38.

［9］Bartelt A,Heeren J.Adipose tissue browning and metabolic health［J］. Nat Rev Endocrinol, 2014,10(1):24-36.

［10］Fedorenko A, Lishko PV, Kirichok Y,et al. Mechanism of fatty-acid-dependent UCP1 uncoupling in brown fat mitochondria［J］.Cell,2012,151 (2): 400-413.

［11］Nedergaard J, Golozoubova V, Matthiaset A,et al. UCPl：the only protein able to mediate adaptive non-shivering tmogenesis and metabolic inefficiency［J］.Biochim Biophys Acta， 2001,1504(1):82-106.

［12］Zuk PA，Zhu M，MorizonoH，et al.Muhilineage cells from human adipose tissue implizations for cells based therapies［J］.Tissue Eng,2001,7(7):211-228.

［13］Dominici M, Le Blanc K, Mueller I,et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement［J］. Cytotherapy,2006,8(4):315-317.

［14］De Ugarte DA, Alfonso Z, Zuk PA,et al. Differential expression of stem cell mobilization-associated molecules on multilineage cells fromadipose tissue and bone marrow［J］. Immunol Lett, 2003,89(2-3):267-270.

［15］Queiroz L, Bonilla L, Malard P,et al. 342 quantitative imaging flow cytometry characterization of functionality and viability of equine adipose-derived stem cells ［J］. Reprod Fertil Deve, 2014,27 (1):259.

［16］Griesche N，Luttmann W，Luttmann A，et al. A simple modification of the separation method reduces heterogeneity of adipose-derived stem cells［J］. Cells Tissues Organs， 2010,192(2):106-115.

［17］Nobusue H，Endo T，Kano K,et al. Establishment of a preadipocyte cell line derived from mature adipocytes of GFP transgenie mice and formation of adipose tissue［J］. Cell Tissue Res,2008,332(3):435-446.

［18］Qi LJ, Guo ZhK, Li Q,et al.Different appearance of senescence and apoptosis during primary culture of the mouse adipose-derived mesenchymal stem cells in vitro［J］. Acta Anatomica Sinica,2012,43(3):366-371.(in Chinese)

齐立杰，郭志坤，李琼, 等，不同年龄小鼠脂肪源间充质干细胞体外原代培养衰老和凋亡的变化［J］.解剖学报，2012,43(3):366-371.

［19］Dimicoli-Salazar S, Bulle F, Yacia A,et al. Efficient in vitro myogenic reprogramming of human primary mesenchymal stem cells and endothelial cells by Myf5［J］. Biol Cell, 2011,103(11):531-542.

［20］Ponrartana S, Aggabao PC, Chavez TA,et al.Changes in brown adipose tissue and muscle development during infancy［J］. J Pediatr, 2016,173:116-121.

［21］Yoshimura K, Suga H, Eto H,et al. Adipose-derived stem/progenitor cells: roles in adipose tissue remodeling and potential use for soft tissue augmentation［J］. Regen Med, 2009,4(2):265-273.

［22］Park E, Patel AN. Changes in the expression pattern of mesenchymal and pluripotent markers in human adipose-derived stem cells［J］. Cell Biol Int, 2010,34(10):979-984.

［23］Nishiwaki S, Nakayama T, Saito S,et al. Efficacy and safety of human adipose tissue-derived mesenchymal stem cells for supporting hematopoiesis［J］. Int J Hematol, 2012,96(3):295-300.

［24］Lin LX, Huang Y, Wang YT,et al. Comparison of biological characteristics of adipose-derived stem cells from different parts［J］. Chinese Journal of Tissue Engineering Research,2013,17(27):4992-4997. (in Chinese)

林立新，黄勇，王玉婷，等. 不同部位脂肪源性干细胞的生物学特性比较［J］. 中国组织工程研究，2013,17(27):4992-4997.

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