Methods comprising apoptosis inhibitors for the generation of transgenic pigs

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Stem Cell Related Patent Number US7101710

Title:Two-stage culture protocol for isolating mesenchymal stem cells from amniotic fluid
Inventors:Tsai, Ming-Song; Taipei, Taiwan Hwang, Shiaw-min; Hsinchu 300, Taiwan
Summary:This invention describes a method for the isolation of mesenchymal stem cells from amniotic fluid via the culturing of amniocytes. Described herein are the means by which mesenchymal stem cells may be harvested from human amniotic fluid via a two-stage culture protocol comprised of culturing the human amniocytes followed by the culturing of mesenchymal stem cells. Further disclosed are methods for preparing the human amniocytes and the primary amniocyte cultures using routine or standard culture protocol in a cytogenetic laboratory, from which the resulting amniotic fluid mesenchymal stem cells may be induced to develop into multi-lineage cells, such as adipocytes, osteocytes, and neuronal cells, among others. Applications are provided in the treatment of heart disease, Parkinson's disease, and diabetes.
Abstract:A method of harvesting mesenchymal stem cells from human amniotic fluid uses a two-stage culture protocol comprising culturing human amniocytes and then culturing mesenchymal stem cells. For culturing human amniocytes, primary amniocyte cultures are set up using routine or standard culture protocol in a cytogenetic laboratory. Non-adherent human amniotic fluids cells in the supernatant medium are collected. For culturing mesenchymal stem cells (“MSC”), the non-adherent cells are centrifuged and then plated with an alpha-modified Minimum Essential Medium supplemented with fetal bovine serum. Incubate with humidified CO2 for MSC growth. Reverse transcription polymerase chain reaction (“RT-PCR”) and immunocytochemical analyses reveal that Oct-4 mRNA and OCT-4 protein expression is detectable in the cultured amniotic fluid mesenchymal stem cells (“AFMSCs”). Under differentiation culture conditions, the AFMSCs can be induced to develop into multi-lineage cells, such as adipocytes, osteocytes, neuronal cells, etc.
US Patent Website:Click Here for Full Text of Patent
Title Number:US7101710
Application Number:US2004000847582
Date Filed:18/05/2004
Date Published:05/09/2006
Assignee:None


 
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