Purpose:To present an experimental model of qualitative and quantitative analysis of mesenchymal stem cells from fat of rabbits obtained by lipectomy. The fat could be a great source for obtaining mesenchymal stem cells and to create conditions for repairing injured tissues by bioengineering. Methods: New Zealand rabbits (n= 10) adipose panicle (2-3 cm) were removed by lipectomy, fragmented and washed with PBS and enzymatically dissociated with trypsin/EDTA. Lately, these cells were incubated in culture medium DMEM and after 20 days, was performed quantitative analysis of the accession of first and second mesenchymal cells in cell culture bottles. Results: The fat total cells (CTF) were 1.62 x10 6 cells/mL and presented 98% of viability. These cells were taken for cultivation and after 20 days were counted 2.88 x10 6 cells/mL MSC. The same was done and after 20 days we quantified 4.28 x10 6 cells/mL MSC. Conclusion: The lipectomy of adipose panicule is a very satisfactory method to extract stem cells from fat, quantitatively and qualitatively. Key words: Lipectomy. Stem Cells. Fats. Rabbits. RESUMOObjetivo: Apresentar um modelo experimental de análise qualitativa e quantitativa de células tronco mesênquimais proveniente da gordura de coelhos obtido por lipectomia. A gordura poderia ser uma grande fonte de obtenção de células tronco mesenquimais, criando condições para a reparação de tecidos lesados. Métodos: Foram removidos os panículos adiposos (2-3 cm) da região cervical de Coelhos Nova Zelândia (n = 10) por lipectomia. Os panículos foram fragmentados e lavados com PBS e, posteriormente, dissociados enzimaticamente com tripsina / EDTA. As células extraídas do panículo adiposo foram incubadas em meio de cultura DMEM e após 20 dias, foi realizada uma análise quantitativa da adesão de primeira e segunda passagem das células mesênquimais em garrafas de cultura. Resultados: Foram extraídas 1,62 x106 cel/ mL células totais de gordura (CTG) with 98% de viabilidade. Essas células foram levadas para o cultivo e após 20 dias, foi realizada a primeira passagem (1pd) sendo quantificadas 2,88 x10 6 cel/mL células tronco mesênquimais (CTM). Na segunda passagem (2pd) foi obtido 4,28 x10 6 cel/mL CTM. Conclusão: A lipectomia do paniculo adiposo é um método muito satisfatório para extrair células tronco a partir de gordura, quantitativamente e qualitativamente. Descritores: Lipectomia. Células-Tronco. Gorduras. Coelhos.

Comparative study of technique to obtain stem cells from bone marrow collection between the iliac crest and the femoral epiphysis in rabbits 1Estudo comparativo da técnica para a obtenção de células tronco pela coleta de medula óssea entre a crista ilíaca e epífise do fêmur em coelhos ABSTRACT Purpose: To assess the technique for the collection of rabbit bone marrow stem cells from different regions to be used as an experimental model in regenerative medicine. Methods: Thirty rabbits were allocated into 2 groups: GROUP A, n=8, animals that underwent bone marrow blood (BMB) harvesting from the iliac crest; and GROUP B: including 22 rabbits that underwent BMB harvesting from the femur epiphysis. After harvesting, mononuclear cells were isolated by density gradient centrifugation (Ficoll -Histopaque). The number of mononuclear cells per ml was counted in a Neubauer chamber and cell viability was checked through Tripan Blue method. Results:Harvesting from the iliac crest yielded an average of 1 ml of BMB and 3,6.10 6 cells/ml over 1 hour of surgery, whereas an average of 3ml of BMB and 11,79.10 6 cells./ml were obtained in 30 min from the femur epiphysis with a reduced animal death rate. Conclusion:The analysis for the obtention of a larger number of mononuclear cells/ml from rabbit bone marrow blood was more satisfactory in the femur epiphysis than in the iliac crest.

Creating experimental models for obtaining stem cells from adipose tissue is necessary to elucidate their peculiar features. Objective: This study proposed a reliable reproducible and consistent experimental model for obtaining mesenchymal stem cells from adipose tissue. Material and Method: Lines of New Zealand rabbits, Wistar rats and CaviaPorcellus guinea pigs (4 animals per species) were used. Fatty tissue mesenchymal stem cells were removed from dorsal, epididymal and inguinal regions. Percentage viable cells and percentage cells expanded and submitted to chondrogenic differentiation were compared by animal species and collection site. Results: Chondrogenic differentiation occurred in a similar manner across all samples, independently of animal species or collection site. Among samples assessed, the inguinal region of rats yielded the highest percentage of viable and expanded cells. Conclusion: A reliable, reproducible and consistent model for obtaining mesenchymal stem cells was produced. Of the several variables analysed, the best results were obtained from the inguinal region of the rat.