PMID- 3165372
OWN - NLM
STAT- MEDLINE
DCOM- 19880913
LR  - 20190828
IS  - 0301-5564 (Print)
IS  - 0301-5564 (Linking)
VI  - 89
IP  - 3
DP  - 1988
TI  - Gradient fractionation of cycling and resting cells monitored by BrdUrd 
      incorporation.
PG  - 241-5
AB  - A number of techniques are currently employed for the fractionation of 
      heterogeneous cell populations or for the separation of cells in different phases 
      of their cycle. With the development of osmotically inert colloidal silica 
      particles media, density gradient centrifugation became an established method for 
      the separation and purification of cells and subcellular particles. We have 
      applied this technique to the separation of cycling from resting Friend 
      erythroleukemia cells, to obtain purified populations for further biological 
      assays. The flow cytometric analysis of DNA content of the different fractions 
      obtained by the gradient and stained with Propidium Iodide (PI), showed the S 
      compartment highly concentrated in the 1.073/77 g/ml interface, while the upper 
      levels of the gradient were highly enriched of cells in G1 phase. Moreover, the 
      dual parameter analysis of DNA content by means of Bromodeoxyuridine (BrdUrd) 
      incorporation and PI staining, showed that part of the cells in the 1.067/73 
      fraction represented the early S phase even if their DNA level, measured on the 
      basis of PI fluorescence was within the diploid cell cluster. This method seems 
      to be suitable to obtain pure cell fractions even when dealing with numerically 
      large populations.
FAU - Papa, S
AU  - Papa S
AD  - Istituto di Anatomia Umana Normale, Universita di Bologna, Italy.
FAU - Vitale, M
AU  - Vitale M
FAU - Mazzotti, G
AU  - Mazzotti G
FAU - Rizzoli, R
AU  - Rizzoli R
FAU - Falconi, M
AU  - Falconi M
FAU - Bartoletti, A
AU  - Bartoletti A
FAU - Manzoli, F A
AU  - Manzoli FA
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Germany
TA  - Histochemistry
JT  - Histochemistry
JID - 0411300
RN  - 0 (DNA, Neoplasm)
RN  - 36015-30-2 (Propidium)
RN  - G34N38R2N1 (Bromodeoxyuridine)
SB  - IM
MH  - Animals
MH  - Bromodeoxyuridine/*metabolism
MH  - *Cell Cycle
MH  - Cell Separation/*methods
MH  - Centrifugation, Density Gradient
MH  - DNA, Neoplasm/metabolism
MH  - Flow Cytometry
MH  - Friend murine leukemia virus
MH  - Leukemia, Erythroblastic, Acute/metabolism/pathology
MH  - Propidium
MH  - Tumor Cells, Cultured/*cytology/metabolism
EDAT- 1988/01/01 00:00
MHDA- 1988/01/01 00:01
CRDT- 1988/01/01 00:00
PHST- 1988/01/01 00:00 [pubmed]
PHST- 1988/01/01 00:01 [medline]
PHST- 1988/01/01 00:00 [entrez]
AID - 10.1007/BF00493147 [doi]
PST - ppublish
SO  - Histochemistry. 1988;89(3):241-5. doi: 10.1007/BF00493147.