Adipose-derived mesenchymal stem cells from liposuction and resected fat are feasible sources for regenerative medicineReportar como inadecuado




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European Journal of Medical Research

, 22:17

Orthopedics

Abstract

BackgroundThe use of mesenchymal stem cells MSCs in research and in regenerative medicine has progressed. Bone marrow as a source has drawbacks because of subsequent morbidities. An easily accessible and valuable source is adipose tissue. This type of tissue contains a high number of MSCs, and obtaining higher quantities of tissue is more feasible. Fat tissue can be harvested using different methods such as liposuction and resection. First, a detailed isolation protocol with complete characterization is described. This also includes highlighting problems and pitfalls. Furthermore, some comparisons of these different harvesting methods exist. However, the later characterization of the cells is conducted poorly in most cases.

MethodsWe performed an in-depth characterization over five passages including an investigation of the effect of freezing and thawing. Characterization was performed using flow cytometry with CD markers, metabolic activity with Alamar Blue, growth potential in between passages, and cytoskeleton staining.

ResultsOur results show that the cells isolated with distinct isolation methods solid versus liposuction -liquid- have the same MSC potential. However, the percentage of cells positive for the markers CD73, CD90, and CD105 is initially quite low. The cells isolated from the liquid fat tissue grow faster at higher passages, and significantly more cells display MSC markers.

ConclusionIn summary, we show a simple and efficient method to isolate adipose-derived mesenchymal stem cells from different preparations. Liposuctions and resection can be used, whereas liposuction has more growth potential at higher passages.

KeywordsAdipose tissue Mesenchymal stem cells Liposuction Solid fat Growth potential Surface characterization AbbreviationsMSCmesenchymal stem cells

CDclusters of differentiation

BMbone marrow

AMSCadipose-derived mesenchymal stem cell

DMEMDulbecco’s modified eagle medium

FBSfetal bovine serum

P-Spenicillin-streptomycin

DPBSDulbecco’s phosphate buffered saline

DMSOdimethyl sulfoxide

RTroom temperature

w-owithout

EDTAethylenediaminetetraacetic acid

CO2carbon dioxide

Sandra Schneider and Marina Unger contributed equally to this work





Autor: Sandra Schneider - Marina Unger - Martijn van Griensven - Elizabeth R. Balmayor

Fuente: https://link.springer.com/







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