In-vitro characterization of serially expanded porcine oesophageal epithelial cells in feeder layer-free culture system for tissue engineering application

The objectives of the study are to isolate and serially expand the porcine oesophageal epithelial cells (PEECs) in serum-free, feeder layer-free culture system, to monitor the growth kinetics in in-vitro cultures and to characterize the stem-like nature, proliferative and differentiation potentia...

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Main Author: Thi, Thi Sein.
Other Authors: Chian Kerm Sin
Format: Theses and Dissertations
Language:English
Published: 2009
Subjects:
Online Access:http://hdl.handle.net/10356/18684
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Institution: Nanyang Technological University
Language: English
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spelling sg-ntu-dr.10356-186842023-03-11T17:33:14Z In-vitro characterization of serially expanded porcine oesophageal epithelial cells in feeder layer-free culture system for tissue engineering application Thi, Thi Sein. Chian Kerm Sin School of Mechanical and Aerospace Engineering DRNTU::Science::Medicine::Tissue engineering DRNTU::Science::Medicine::Biomedical engineering The objectives of the study are to isolate and serially expand the porcine oesophageal epithelial cells (PEECs) in serum-free, feeder layer-free culture system, to monitor the growth kinetics in in-vitro cultures and to characterize the stem-like nature, proliferative and differentiation potential of cultured PEECs in-vitro with the use of immunocytochemical method. The porcine oesophageal epithelial cells were isolated by two-step enzymatic methods using dispase digestion at physiological temperature for earlier establishment of primary cultures. The primary cultures of PEECs were established from porcine oesophageal tissue collected directly from the abattoir. The cells could be cultured in serum-free, feeder layer-free culture medium with reasonable proliferation up to passage 6 – 8 although optimization of culture medium was needed for extended in-vitro growth. Sorting the cells depending on the rapid adhesiveness to extracellular matrix proteins coated on the culture surface could enrich highly proliferative cells with extended culture lifespan in-vitro. The morphology of the cultured cells monitored under light microscope confirmed their morphological stability in in-vitro cultures. The growth kinetics of the cultured cells was assessed by population doubling time and serial MTS assays. Master of Science (Biomedical Engineering) 2009-07-02T06:41:14Z 2009-07-02T06:41:14Z 2009 2009 Thesis http://hdl.handle.net/10356/18684 en 116 p. application/pdf
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic DRNTU::Science::Medicine::Tissue engineering
DRNTU::Science::Medicine::Biomedical engineering
spellingShingle DRNTU::Science::Medicine::Tissue engineering
DRNTU::Science::Medicine::Biomedical engineering
Thi, Thi Sein.
In-vitro characterization of serially expanded porcine oesophageal epithelial cells in feeder layer-free culture system for tissue engineering application
description The objectives of the study are to isolate and serially expand the porcine oesophageal epithelial cells (PEECs) in serum-free, feeder layer-free culture system, to monitor the growth kinetics in in-vitro cultures and to characterize the stem-like nature, proliferative and differentiation potential of cultured PEECs in-vitro with the use of immunocytochemical method. The porcine oesophageal epithelial cells were isolated by two-step enzymatic methods using dispase digestion at physiological temperature for earlier establishment of primary cultures. The primary cultures of PEECs were established from porcine oesophageal tissue collected directly from the abattoir. The cells could be cultured in serum-free, feeder layer-free culture medium with reasonable proliferation up to passage 6 – 8 although optimization of culture medium was needed for extended in-vitro growth. Sorting the cells depending on the rapid adhesiveness to extracellular matrix proteins coated on the culture surface could enrich highly proliferative cells with extended culture lifespan in-vitro. The morphology of the cultured cells monitored under light microscope confirmed their morphological stability in in-vitro cultures. The growth kinetics of the cultured cells was assessed by population doubling time and serial MTS assays.
author2 Chian Kerm Sin
author_facet Chian Kerm Sin
Thi, Thi Sein.
format Theses and Dissertations
author Thi, Thi Sein.
author_sort Thi, Thi Sein.
title In-vitro characterization of serially expanded porcine oesophageal epithelial cells in feeder layer-free culture system for tissue engineering application
title_short In-vitro characterization of serially expanded porcine oesophageal epithelial cells in feeder layer-free culture system for tissue engineering application
title_full In-vitro characterization of serially expanded porcine oesophageal epithelial cells in feeder layer-free culture system for tissue engineering application
title_fullStr In-vitro characterization of serially expanded porcine oesophageal epithelial cells in feeder layer-free culture system for tissue engineering application
title_full_unstemmed In-vitro characterization of serially expanded porcine oesophageal epithelial cells in feeder layer-free culture system for tissue engineering application
title_sort in-vitro characterization of serially expanded porcine oesophageal epithelial cells in feeder layer-free culture system for tissue engineering application
publishDate 2009
url http://hdl.handle.net/10356/18684
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