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Optimization of Buffalo (Bubalus bubalis) Embryonic Stem Cell Culture System
Mohammad Zandi1, Musharifa Muzaffar2, Syed Mohmad Shah2
1Department of Animal and Poultry Science and Fisheries, Agricultural Institute, Iranian Research Organisation for Science and Technology (IROST), Tehran, Iran.
Cell Journal
|July 23, 2015
Summary
Developing feeder-free buffalo embryonic stem cell (ES cell) cultures is crucial for advancing stem cell research. This study explores methods to maintain ES cell pluripotency without feeder layers, enabling more reliable experimental outcomes.
Area of Science:
- Stem Cell Biology
- Reproductive Biology
- Animal Biotechnology
Background:
- Embryonic stem (ES) cells require feeder cell layers to maintain pluripotency.
- Feeder layers complicate research by introducing confounding variables.
- Developing feeder-free systems is essential for advancing stem cell research.
Purpose of the Study:
- To establish and characterize a buffalo ES cell line.
- To develop feeder-free culture systems for buffalo ES cells.
- To evaluate factors supporting buffalo ES cell growth and pluripotency.
Main Methods:
- Established buffalo ES cell line from in vitro blastocysts.
- Characterized pluripotency using Alkaline phosphatase and immunofluorescence.
- Tested fibroblast growth factor 2 (FGF-2), leukemia inhibitory factor (LIF), and Y-27632 in feeder-free cultures.
- Assessed feeder-conditioned media effects on gelatin-coated cultures.
Main Results:
- Y-27632 with FGF-2 and LIF enhanced colony growth and Nanog expression.
- Feeder-conditioned media improved buffalo ES cell growth on gelatin.
- Feeder layer cultures supported buffalo ES cells for over two years.
- Feeder-free systems provide short-term maintenance, while feeder layers ensure long-term pluripotency.
Conclusions:
- A feeder-free culture system was developed for short-term maintenance of buffalo ES cells.
- Feeder layer-based cultures support long-term maintenance of buffalo ES cell pluripotency.
- These findings contribute to optimizing stem cell culture techniques in buffalo.
