Abstract
BackgroundAdult stem cells appear to be a promising subject for tissue engineering, representing an individual material for regeneration of aged and damaged cells. Especially adipose derived stromal cells (ADSC), which are easily to achieve, allow an encouraging perspective due to their capability of differentiating into miscellaneous cell types. Here we describe the in vitro formation of human subcutaneous, visceral and omental ADSC micromasses and compare their histological attributes while being cultivated on collagen membranes.MethodsSubcutaneous, visceral and omental fat tissue derived cells were isolated and processed according to standard protocols. Positively stained cells for CD13, CD44 and CD90 were cultivated on agarose in order to study micromass formation using a special method of cell tracking. Stained paraffin-embedded micromasses were analysed morphologically before and after being plated on collagen membranes.ResultsThe micromass formation process was similar in all three tissue types. Subcutaneous fat tissue derived micromasses turned out to develop a more homogeneous and compact shape than visceral and omental tissue. Nevertheless all micromasses adhered to collagen membranes with visible spreading of cells. The immune histochemical (IHC) staining of subcutaneous, visceral and omental ADSC micromasses shows a constant expression of CD13 and a decrease of CD44 and CD 90 expression within 28 days. After that period, omental fat cells don’t show any expression of CD44.ConclusionIn conclusion micromass formation and cultivation of all analysed fat tissues can be achieved, subcutaneous cells appearing to be the best material for regenerative concepts.
Highlights
Adult stem cells appear to be a promising subject for tissue engineering, representing an individual material for regeneration of aged and damaged cells
Micromass formation of subcutaneous, visceral and omental fat tissue derived cells was displayed by the software “MicroTrac”
We showed that the morphology of subcutaneous, visceral and omental fat tissue derived micromasses differs in cell density and the inclusion of other cell types
Summary
Adult stem cells appear to be a promising subject for tissue engineering, representing an individual material for regeneration of aged and damaged cells. Adipose derived stromal cells (ADSC), which are to achieve, allow an encouraging perspective due to their capability of differentiating into miscellaneous cell types. Adipose derived stromal cells (ADSC) are multipotent cells well known in the literature. They occur in abundance, are to achieve and able to differentiate into miscellaneous lineages like osteoblasts, endothelial cells, or neurogenic cells without difficulties. For further clinical use of the cells it was relevant to know whether all the isolated cell types are able to form micromasses, three-dimensional cell cultures. Without an anchoring material like the bottom of a petri dish, cells use to congregate in order to form a so-called sphere, which displays both an in vitro tissue-model and a circumscribed three-dimensional source of tissue regeneration. In micromasses of hADSC, a significant increase of angiogenic growth factors, as HGF, VEFG and FGF2, was found in comparison to a monolayer culture. [3] a strong influence on their differentiation
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