Human amnion as a novel cell delivery vehicle for chondrogenic mesenchymal stem cells
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Human amnion as a novel cell delivery vehicle for chondrogenic mesenchymal stem cells Sik-Loo Tan • Sofiah Sulaiman • Belinda Pingguan-Murphy • L. Selvaratnam Cheh-Chin Tai • T. Kamarul
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Received: 10 October 2008 / Accepted: 17 November 2009 / Published online: 2 December 2009 Ó Springer Science+Business Media B.V. 2009
Abstract This study investigates the feasibility of processed human amnion (HAM) as a substrate for chondrogenic differentiation of mesenchymal stem cells (MSCs). HAM preparations processed by air drying (AD) and freeze drying (FD) underwent histological examination and MSC seeding in chondrogenic medium for 15 days. Monolayer cultures were used as control for chondrogenic differentiation and HAMs without cell seeding were used as negative control. Qualitative observations were made using scanning electron microscopy analysis and quantitative
analyses were based on the sulfated glycosaminoglycans (GAG) assays performed on day 1 and day 15. Histological examination of HAM substrates before seeding revealed a smooth surface in AD substrates, while the FD substrates exhibited a porous surface. Cell attachment to AD and FD substrates on day 15 was qualitatively comparable. GAG were significantly highly expressed in cells seeded on FD HAM substrates. This study indicates that processed HAM is a potentially valuable material as a cell-carrier for MSC differentiation. Keywords Cell delivery vehicle Chondrogenic differentiation Human amnion Mesenchymal stem cells
S.-L. Tan C.-C. Tai T. Kamarul (&) Department of Orthopaedic Surgery, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia e-mail: [email protected]; [email protected] S. Sulaiman Department of Obstetric and Gynaecology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia B. Pingguan-Murphy Department of Biomedical Engineering, Faculty of Engineering, University of Malaya, 50603 Kuala Lumpur, Malaysia L. Selvaratnam School of Medicine and Health Sciences, Monash University, Sunway Campus, 46150 Subang Jaya, Selangor, Malaysia
Introduction Surgical application of the human amnion (HAM) was first introduced in the early twentieth century as a novel and highly effective means for treating open wounds and burns (Davis 1910). It was only in later years that its potential was realized, resulting in a number of clinical applications being successfully introduced, including, for use in ophthalmological, gynaecological, urological and neurological surgery (Quinby et al. 1982; Subramanyam 1995; Ward et al. 1989). HAM itself is the innermost layer of the human placenta comprising a single layer of non-ciliated
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cuboidal cells. It is a tough, alymphatic, avascular and aneural tissue with a shiny physical appearance (Mohamad 2001) and has been reported to posses antimicrobial properties, which include impermeability to a diverse panel of bacteria (Kjaergaard et al. 2001; Singh et al. 2007). Recent research has expanded the use of HAM into a new area in tissue engineering. Demonstration of its use
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