Frozen adipose-derived mesenchymal stem cells maintain high capability to grow and differentiate.

In recent years, there has been a shift toward tissue-engineering strategies using stem cells for plastic and reconstructive surgical procedures. Therefore, it is important to develop safe and reproducible protocols for the extraction of adipose-derived stromal cells (ASCs) to allow cells to be stored in liquid nitrogen for future needs. The aspirated liposuction obtained from healthy donors were immediately processed after the suction using a protocol developed in our laboratory. The resulting stromal vascular fraction (SVF) was then characterized by the presence of adipose-derived stromal cells, at later stage frozen in liquid nitrogen. After that, cells were thawed and again characterized by adipose-derived stromal cells, cellular survival, differentiation ability and Colony Forming Unit-Fibroblast like colonies (CFU-F). Extraction and freezing of cells contained in the stromal vascular fraction demonstrate that thawed cells maintain the full capability to grow and differentiate in culture. The advent of adipose-derived stromal cells use in tissue engineering will assume a wide role in esthetic restoration in plastic surgery. It is thus important to develop clinically translatable protocols for the preparation and storage of adipose-derived stromal cells. Our results show that adipose-derived stromal cells in serum free can easily be frozen and stored in liquid nitrogen with retention of 85% of cell viability and 180,890 cell/g yield plus normal proliferative capacity and differentiation potential compared with fresh controls. These observations set the basis for adipose-derived stromal cells banking.

Phonetic and otological results after early palate closure in 18 consecutive children presenting with cleft lip and palate.

OBJECTIVE: First, to analyze the speech and hearing results at 3.5 years of age when early palate closure has been performed. Second to assess at 7 years of age the need for speech therapy and/or additional surgery in the form of cranial-based pharyngeal flap for obtaining normal speech. METHODS: Retrospective study in a tertiary teaching hospital concerning 18 consecutive cases presenting cleft lip and palate with no associated abnormalities. Interventions included early palatine closure (velum at 3 months, hard palate and lip at 6 months). Speech therapy was initiated at 3.5 years of age when needed. Cranial-based pharyngeal flap was performed when normal nasal emission was not obtained by speech therapy at 7 years of age. Phonetic and otological assessment were performed in all cases. RESULTS: Good to excellent speech in the majority (95%) of children, with only 3/18 undergoing pharyngoplasty to obtain type I or I/II speech by the age of 7 (range: 6.5-8.0). 6/18 children had drain insertion, and 2/18 had hearing loss of 20-40 dB in all frequencies. CONCLUSION: Most children (95%) start school with good or excellent speech. However, the high incidence of middle ear problems shows that more effective screening and treatment are warranted.