Cleft lip and palate is a congenital malformation that requires a multidisciplinary treatment that evolves pediatrician, obstetrics, fetal medicine, genetics, plastic surgery, orthodontics, speech therapist, nursery, and psychology. Actually, the authors believe that it could be possible to ad protocols to use stem cells.The intrauterine diagnosis leads to preborn parental orientation and better parental collaboration to accept a precocious multidisciplinary treatment. After birth the authors' protocol is: orthodontic devices, phonoaudiology, and surgical procedures.The authors' cleft lip and palate reconstructive surgery protocol demands several steps and begins at 4 to 6-month old with rhinocheiloplasty and soft palate closure at the same moment. The treatment sequence involves the hard palate surgery (8-18 months after the first surgical step), alveoloplasty (after 10 years old), and secondary rhinoplasty (after 14 years old).New ideas to use stem cells and blood from the umbilical cord and also blood from placenta are discussed to improve final surgical results. Maternal stem cells are easy to collect, there are no damage to the patient and mother, it is autologous and it could be very useful in the authors' protocol.Nine patients with clef lip and palate were operated and had stem cells from umbilical cord blood and placenta blood injected into the bone and soft tissue during the primary procedure (rhinocheiloplasty).The stem cells activity into soft tissue and bone were evaluated. Preliminary results have shown no adverse results and improvement at the inflammatory response. A treatment protocol with stem cells was developed. It had a long time follow-up of 10 years.

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.

Injection of skin fibroblasts cultivated in medium supplemented with human serum is a viable technique and provokes no side effects. Four injections given at 15-day intervals containing a total of 6.4 × 10(6) fibroblasts/mL resulted in significant improvement in periorbital skin flaccidity. Further studies should be conducted with larger sample sizes.

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