Type

Journal Article

Authors

Lorraine O'Driscoll
Martin Clynes
Sweta Rani

Subjects

Biochemistry

Topics
animals preproinsulin insulin secreting cells deoxyribonucleases glucose cell function protein precursors mice rna messenger extracellular space culture media conditioned mass cell line reverse transcriptase polymerase chain reaction cells cell count detection cytology humans secretion insulin beta cell chemistry ribonucleases genetics cercopithecus aethiops metabolism proinsulin mrna

Detection of amplifiable mRNA extracellular to insulin-producing cells: potential for predicting beta cell mass and function. (2007)

Abstract Detecting extracellular nucleic acids in the serum/plasma of cancer patients may help in cancer diagnosis. We investigated whether extracellular mRNAs are reproducibly detectable in conditioned medium (CM) from insulin-producing cell cultures and if their presence and amounts are indicative of cell number and/or function. We isolated mRNA from medium conditioned by the culture of several insulin-producing cell types: MIN6(L) (glucose-responsive), MIN6(H) (glucose-nonresponsive), and MIN6 B1 murine beta cells and monkey kidney fibroblast cells engineered to produce human preproinsulin (PPI) (Vero-PPI). We used reverse transcription-PCR analyses to evaluate the occurrence of several mRNAs and investigated whether the presence and amounts of the various extracellular mRNAs are associated with cell mass and/or function. Reproducible amplification of mRNAs encoded by Pdx1, Npy, Egr1, Pld1, Chgb, Ins1, Ins2, and Actb from MIN6(L), MIN6(H), and MIN6 B1 cells and their CM suggests that beta cells transcribe and release these mRNAs into their culture environment. Similarly, PPI mRNA was detected in samples of Vero-PPI cells and CM. The amounts of some mRNAs reflected the numbers and functional status (i.e., glucose responsiveness vs nonresponsiveness) of the cells conditioning the medium. Although Pax4 mRNA was detected in the MIN6 B1 cell line, the fact that this transcript was not amplifiable from the corresponding CM suggested that mRNA release was selective. mRNAs may be secreted from insulin-producing cells, are reproducibly detected in the extracellular environment, and may have potential as extracellular biomarkers for assessing beta cell mass and function.
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Full list of authors on original publication

Lorraine O'Driscoll, Martin Clynes, Sweta Rani

Experts in our system

1
Lorraine O'Driscoll
Trinity College Dublin
Total Publications: 152
 
2
Martin Clynes
Dublin City University
Total Publications: 209
 
3
Sweta Rani
Trinity College Dublin
Total Publications: 22