Post-culture treatment protocols for PLGA membrane scaffolds

Marianne J Ellis, R Forsey, J B Chaudhuri

Research output: Contribution to journalArticle

Abstract

The interactions of post-culture treatments reagents used for fixing, lysing and cell quantification on poly(lactide-co-glycolide) (PLGA) flat sheet membrane scaffolds are presented. Lysing with Alkaline buffer solution/Triton X-100/MilliQ water (ATM) and fixing with 10% Neutral Buffered Formalin (10% NBF) had no affect on membrane structure while fixing with 95% ethanol caused smoothing of the surface, shrinkage and a reduction in surface area of 55, 48 and 33, for 100:0, 75:25 and 50:50 (PLA:PGA), respectively. PicoGreen assay was selected for cell (560pZIPv.neo) quantification since the background noise would not affect readings for cell numbers over 3,000 cells/cm(2), while the background reading was too high for MTT and Methylene Blue (MB). MB at 0.5% (w/v) was, however, deemed suitable for visualising cell morphology on the membranes. Furthermore ATM buffer was suitable for the PicoGreen assay, which allows the same samples to be used for quantification of alkaline phosphatase activity.
Original languageEnglish
Pages (from-to)215-222
Number of pages8
JournalBiotechnology Letters
Volume32
Issue number2
Early online date10 Oct 2009
DOIs
Publication statusPublished - 1 Feb 2010

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Clinical Protocols
Membranes
Methylene Blue
Reading
Buffers
Prostaglandins A
Polyglactin 910
Octoxynol
Formaldehyde
Alkaline Phosphatase
Noise
Ethanol
Cell Count
polylactic acid-polyglycolic acid copolymer
Water
PicoGreen
Therapeutics

Cite this

Post-culture treatment protocols for PLGA membrane scaffolds. / Ellis, Marianne J; Forsey, R; Chaudhuri, J B.

In: Biotechnology Letters, Vol. 32, No. 2, 01.02.2010, p. 215-222.

Research output: Contribution to journalArticle

Ellis, Marianne J ; Forsey, R ; Chaudhuri, J B. / Post-culture treatment protocols for PLGA membrane scaffolds. In: Biotechnology Letters. 2010 ; Vol. 32, No. 2. pp. 215-222.
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