Development of a procedure to simultaneously isolate RNA, DNA, and proteins for [corrected] characterizing cells invading or cultured on chitosan scaffolds.
For many years, chitosan and its derivatives have been considered to be promising biomaterials for tissue engineering and repair. However, information regarding their biological effect on cell phenotype is usually limited to evaluation of cell proliferation and survival, overlooking proteomic and transcriptomic analysis. This is largely related to the lack of efficient and quantitative procedures for protein and nucleic acid purification from cells cultured on, or inside, chitosan scaffold. Here we describe an ultracentrifugation procedure enabling the simultaneous and quantitative recovery of high quality RNA, DNA and proteins from cells growing in close contact of biomaterial matrices containing chitosan.
Tchemtchoua
Victor T
VT
Laboratory of Connective Tissues Biology, GIGA Research Center, University of Liège, 4000 Sart Tilman, Belgium.
Atanasova
Ganka
G
Aqil
Abdelhafid
A
Maquet
Véronique
V
Jerôme
Christine
C
Poumay
Yves
Y
Colige
Alain
A
D000818:Animals; D001671:Biochemistry / Q000379:methods*; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D048271:Chitosan; D004247:DNA / Q000302:isolation & purification*; D051379:Mice; D011506:Proteins / Q000302:isolation & purification*; D012313:RNA / Q000302:isolation & purification*
D011506:Proteins; D012313:RNA; D004247:DNA; D048271:Chitosan