| Objective:Collect the terminal hardness tumor tissue at the end of tumescent lipoma liposuction procedure and get lipomas through surgical excision, collect adipose tissue around lipomas. Makers of the adipose stem Cells (ASCs) were used to sign ASCs, to explore the role of recurrence of terminal hardness tumor tissue afteroperation of lipoma liposuction. Methods:1 The specimens were collected from 2 multiple lipomatosis male patients, 28 lipomas that diameter of lipomas between 2.5cm-4.5cm were selected for lipoma liposuction. 3 lipomas were removed completely, adipose tissue size 0.5cm around lipoma was collected, the total of specimens was 18. All specimens were fixed in 4% formalin solution for 72 hours; specimens were paraffin-embed after dehydration and trimming.2 Slice the specimens into section with 8μm thickness, routine HE stained was executed,observed by light microscope and pictures captured,difference of three sorts specimens was evaluated .3 Slice the specimens into section with 8μm thickness, immunohistochemical stain with antibody of maker of adipose stem cell CD29, CD34 and CD44, nucleus afterstained by hematoxylin, dehydration and mounting, observed by light microscope and pictures of positive cells of 2 sorts specimens captured were observed and evaluated.4 Independence-samples T test on density of positive cells; area of positive cells was estimated with SPSS16.0 system,the test standardα=0.05.Results:1 Macropathology The appearance of terminal hardness tumor tissue is primrose, moderate hardness conglomeration , there is floppy, irregular,gray and cord-like around it(Fig.2).2 Abundant stromas exist in terminal hardness tumor tissue, spindle-shaped, fibroblast-like cells located among adipocytes irregularly, nucleus is large and blue stained, cytoplasm is few; differentiation of cells of adipose tissue is mature, fat vacuoles were left and the nucleus located periphery near the plasmalemma , less spindle-shaped, fibroblast-like cells located among adipocytes were observed(Fig.7-8).3 The slices were stained with antibody of maker (CD29, CD34, CD44) of stem cell by Immunohistochemical, plasmalemma and cytoplasm of the spindle-shaped, fibroblast-like cells were stained in brown; it is confirmed that spindle-shaped, fibroblast-like cells are adipose stem cell(Fig. 9-14).4 The density and area of positive cells was analyzed by SPSS16.0 system.4.1 The average density of CD29 stained cells in terminal hardness tumor tissue, adipose tissue by immunohistochemical image analysis is as follows: 410±177cells/mm2 and 74±17cells/mm2; The area of positive cells is as follows: (1.47±0.72)×10-2mm2 and (0.42±0.07)×10-2mm2.4.2 The average density of CD34 in terminal hardness tumor tissue, lipoma and adipose tissue through immunohistochemical image analysis is as follows: 436±157cells/mm2 and 136±31cells/mm2; the area of positive cells is as follows: (1.53±0.68)×10-2mm2 and (0.59±0.10)×10-2mm2.4.3 The average density of CD44 in terminal hardness tumor tissue and adipose tissue through immunohistochemical image analysis is as follows:468±200 cells/mm2 and 80±17cells/mm2; the area of positive cells is as follows: (1.49±0.73)×10-2mm2 and (0.525±0.18)×10-2mm2.5 With SPSS16.0 system, Independence-samples T test on density and area of positive cells were estimated.5.1 The parameter of density analysis of each group, Independence-samples T test display : t'CD29 =9.936 , P<0.01 ; t'CD34=9.787 , P<0.01 , t'CD44=10.17,P<0.01, difference significantly between groups inα=0.05 level.5.2 The parameter of area analysis of each group, Independence-samples T test display: t'CD29=7.61,P<0.05; t'CD34=11.706,P<0.01; t'CD44=6.641, P<0.01, difference significantly between groups inα=0.05 level.Conclusion:1 Abundant adipose stem cells (ASCs) that signed by antibody of stem cell maker CD29, CD34 and CD44 by immunohistochemical stain exists in terminal hardness tumor tissue,2 The density and area of ASCs is much more in terminal hardness tumor tissue than that adipose tissue, the difference is significantly.3 Abundant fibroblast-like cells that signed by the marker of adipose stem cells exist in terminal hardness tumor tissue, which probably is one of the factors lead to lipoma recurrence.
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