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Revista Salud Uninorte
Print version ISSN 0120-5552On-line version ISSN 2011-7531
Abstract
CELIS, Luis Gustavo et al. Isolation and identification of stem cells derived of infrapatelar fat pad. Salud, Barranquilla [online]. 2009, vol.25, n.2, pp.189-196. ISSN 0120-5552.
Introduction: The infrapatellar fat pad (sometimes know Hoffa's pad) is a soft tissue that lies beneath the patella (kneecap) separating it from the femoral condyle. There is evidence suggesting the presence of stem cells on adipose tissue in the adult human. Objective: The purpose of this work is to isolate and identify grown stem cells from Hoffa's fat samples obtained from patients undergoing surgeries exposing this tissue. Materials and Methods: After obtaining informed consent, the biopsy of Hoffa's fat pad was obtained. The samples were incubated with collagenase and PBS a 37 °C and agitated, then it was inactivated with fetal bovine serum and centrifuged, washed twice with PBS, the pellet was resuspended and one part was cultivated on Mesencult medium the other part was used for flow cytometry and stained with methylene blue for morphologic analysis. Also before and after the enzyme digestion, the samples were added to 10% formaldehyde to evaluate the collagenase treatment. Results and analysis: The results show the effectiveness of the enzymatic treatment, the architecture of the adipose tissue was lost. The star shape stem cells morphology was appreciated with methylene blue in the cultures, it was corroborated by flow cytometry with CD13, CD29, CD59 and CD 105 markers. Conclusions: Primary cultures are consolidating, the next aim will be to obtain differentiated specific cell types that can be use in the study of obesity, diabetes and articular illnesses.
Keywords : Stem cells; infrapatellar fat pad; mandis cells; culture cellular.