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Expansion and Osteogenic Differentiation of Human Amniotic Fluid Derived Stem Cells

Abstract Details

2013, Doctor of Philosophy, Ohio State University, Chemical and Biomolecular Engineering.
Because of their easy accessibility and broad multipotentiality, human amniotic fluid stem cells (hAFSCs) are emerging as an important cell source for tissue engineering and regenerative medicine. However, due to the very recent identification of hAFSCs, few research exploring their mass production, differentiation and medium optimization have been carried out. To date, hAFSC cultures are still performed in cell culture dishes, T-flasks and multiwell plates. These traditional techniques exhibit several serious drawbacks, including being labor-intensive, time-consuming, expensive, prone to contamination, and difficult to scale up. For clinical applications, a large number of hAFSCs are required, and conventional cell culture systems cannot meet this demand. In this study, a three-dimensional (3-D) polyethylene terephthalate (PET) fibrous bed bioreactor (FBB) was developed for hAFSC expansion. This novel bioreactor provide large specific surface areas favoring hAFSC attachment and proliferation, better mimic in vivo environments, facilitate nutrient and oxygen diffusion, and protect cells from shear damage. It achieved 155 expansion fold production of functional hAFSCs. Bone diseases, especially osteoporosis, leading to a high risk of fracture and deformities, bring serious issues to public health. Estrogen replacement therapy and biphosphonate have been considered to be the most effective treatment for osteoporosis in the past 10 years. However, they could increase the risk of cancer and cardiovascular diseases and cause acute incapacitating bone, joint, and muscle pain. AFSCs provide a novel cell source for cell therapy, and are reported to have strong potential to differentiate into osteoblasts and chondrocytes. Recently, some medical plant-derived estrogen-like chemical compounds have been shown to have antiosteoporotic activity and minimal side effects. In this study, two natural plant ingredients, naringin and curculigoside, were investigated for their stimulation effects on proliferation and osteogenic differentiation of hAFSCs. The results demonstrated that these two natural ingredients could promote the proliferation and osteogenesis, and concurrently inhibit osteoclastogenesis of hAFSCs. Moreover, signal transduction pathways underlying the promotion of osteogenic differentiation of hAFSCs were clearly revealed. Being a recently identified stem cell for promising clinical applications, hAFSCs have not been well studied for their in vitro expansion. To date, very few medium formulae have been optimized for growth and expansion of hAFSCs. Currently, most hAFSC cultures are carried out in a commonly used serum-rich medium, which is not qualified for good manufacturing practice (GMP). In this study, by cooperating with scientists at Irvine Scientific, the author evaluated three newly developed and optimized media, including a complete medium and two serum-free media, for supporting in vitro proliferation of hAFSCs. The results indicated that the serum-containing complete (SCC) medium could support a faster proliferation than the control medium while maintaining the characteristic immunophenotype and the multilineage differentiation potential of hAFSCs. However, the two serum-free media SF I and SF II were not satisfactory and further development and optimization are needed. Nevertheless, this study provided groundwork for the development of medium for the expansion of hAFSCs for clinical applications.
Shang-Tian Yang (Advisor)
Jeffrey Chalmers (Committee Member)
Andre Palmer (Committee Member)
241 p.

Recommended Citations

Citations

  • Liu, M. (2013). Expansion and Osteogenic Differentiation of Human Amniotic Fluid Derived Stem Cells [Doctoral dissertation, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1370277490

    APA Style (7th edition)

  • Liu, Meimei. Expansion and Osteogenic Differentiation of Human Amniotic Fluid Derived Stem Cells . 2013. Ohio State University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=osu1370277490.

    MLA Style (8th edition)

  • Liu, Meimei. "Expansion and Osteogenic Differentiation of Human Amniotic Fluid Derived Stem Cells ." Doctoral dissertation, Ohio State University, 2013. http://rave.ohiolink.edu/etdc/view?acc_num=osu1370277490

    Chicago Manual of Style (17th edition)