PMID- 3618227
OWN - NLM
STAT- MEDLINE
DCOM- 19870921
LR  - 20191029
IS  - 0108-0164 (Print)
IS  - 0108-0164 (Linking)
VI  - 95
IP  - 4
DP  - 1987 Jul
TI  - Flow cytofluorometry of lysosomal acridine orange uptake by living cultured 
      cells. Effect of trypsinization and starvation.
PG  - 159-65
AB  - The uptake of the fluorescent, lysosomotropic weak base acridine orange (AO) by 
      living cells in culture was studied by flow cytofluorometry. A mouse myeloma cell 
      line (SP 2/0), growing in suspension, and an anchorage-dependent human malignant 
      glioma cell line (U-251 MG), brought into suspension by trypsinization, were 
      used. The consequences of trypsinization were also studied using static 
      cytofluorometry. The lysosomal accumulation of AO by myeloma cells growing in 
      suspension was found to be only moderately affected by starvation (i.e. 
      incubation without medium change) for a period of up to five days. Trypsinization 
      of the glioma cells after staining with AO caused pronounced release of the 
      fluorescent dye while trypsinization before staining with AO did not 
      significantly change the average lysosomal concentration of AO. We did, however, 
      notice certain side effects of trypsinization in the form of both increased 
      cellular green fluorescence and greater intercellular variability that reduce the 
      validity of data obtained from cells detached by routine trypsinization. In 
      conclusion, the condition of the lysosomal vacuome of living cultured cells 
      growing in suspension may be studied by flow cytofluorometry after vital staining 
      with the lysosomotropic weak base AO. Anchorage-dependent trypsinized cells, 
      however, yield unsatisfactory results when examined in a flow cytofluorometer 
      system and are better studied while still attached to their substratum, using 
      static cytofluorometry.
FAU - Olsson, M
AU  - Olsson M
FAU - Rundquist, I
AU  - Rundquist I
FAU - Brunk, U
AU  - Brunk U
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Denmark
TA  - Acta Pathol Microbiol Immunol Scand A
JT  - Acta pathologica, microbiologica, et immunologica Scandinavica. Section A, 
      Pathology
JID - 8206622
RN  - EC 3.4.21.4 (Trypsin)
RN  - F30N4O6XVV (Acridine Orange)
SB  - IM
MH  - Acridine Orange/*metabolism
MH  - Animals
MH  - Cells, Cultured
MH  - *Flow Cytometry
MH  - Glioma/metabolism
MH  - Humans
MH  - Hydrogen-Ion Concentration
MH  - Lysosomes/*metabolism
MH  - Mice
MH  - Multiple Myeloma/metabolism
MH  - Trypsin/*pharmacology
EDAT- 1987/07/01 00:00
MHDA- 1987/07/01 00:01
CRDT- 1987/07/01 00:00
PHST- 1987/07/01 00:00 [pubmed]
PHST- 1987/07/01 00:01 [medline]
PHST- 1987/07/01 00:00 [entrez]
AID - 10.1111/j.1699-0463.1987.tb00025_95a.x [doi]
PST - ppublish
SO  - Acta Pathol Microbiol Immunol Scand A. 1987 Jul;95(4):159-65. doi: 
      10.1111/j.1699-0463.1987.tb00025_95a.x.