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Updated: Jun 23, 2026
Differentiation and Characterization of Neural Progenitors and Neurons from Mouse Embryonic Stem Cells
Published on: May 15, 2020
Rat bone marrow derived mesenchymal progenitor cells support mouse ES cell growth and germ-like cell differentiation
Guanghui Cui1, Zhengyu Qi, Xin Guo
1Medical Center of Peking University & Hong Kong Science and Technology University, Guangdong, China.
Abstract:
Mouse embryonic fibroblasts (MEFs) have been used as feeder cells to support the growth of mouse embryonic stem cell (mESC) and primordial germ cells (PGC) in culture for many years. However, MEF preparation is a complex and tedious task. Recently, there are reports indicating that the microenvironment provided by bone marrow stromal cells could support the survival of embryonic-like stem cells in bone marrow. In this report, rat bone marrow derived mesenchymal progenitor cells (MPC) were used as feeder cells to culture mouse Oct4-GFP ES cell and ES cell derived germ cells. FACS results show that similar to MEF, rat MPC could efficiently support growth of the mouse Oct4-GFP ES cell line in culture (MPC 85.5 +/- 5.1% vs MEF 84.1 +/- 6.2%). ES cells could be subcultured for >15 passages without losing morphological characteristics. The cultured cells expressed stem cell marker alkaline phosphatase, Oct4, Sox2, and SSEA-1. Furthermore, rat MPC cells were able to support survival of germ cells isolated from mouse Oct4-GFP ES cell formed embryoid bodies (EB). After induction by retinoic acid for 7 days, some isolated cells differentiated to spermatogonial stem-like cells, expressing Mvh, Stra-8, Hsp90-a, integrinb1 and a6. Compared with traditional MEF culture systems, the rat MPC culture system is effective in supporting ES cell growth and is easy to prepare.
Insights
Rat bone marrow mesenchymal progenitor cells (MPC) effectively support mouse embryonic stem cell (mESC) growth, offering an easier alternative to traditional mouse embryonic fibroblasts (MEF) feeder layers.
Area of Science:
- Stem Cell Biology
- Developmental Biology
- Cell Culture Technology
Background:
- Mouse embryonic fibroblasts (MEFs) are standard feeder cells for culturing mouse embryonic stem cells (mESCs) and primordial germ cells (PGCs).
- MEF preparation is labor-intensive and complex.
- Bone marrow stromal cells show potential for supporting embryonic-like stem cells.
Purpose of the Study:
- To evaluate rat bone marrow-derived mesenchymal progenitor cells (MPCs) as an alternative feeder layer for culturing mouse ESCs and germ cells.
- To compare the efficacy of MPCs with traditional MEFs for stem cell culture.
Main Methods:
- Culture of mouse Oct4-GFP ESCs and ESC-derived germ cells on rat MPC feeder layers.
- Flow cytometry analysis (FACS) to assess cell growth and viability.
- Assessment of stem cell marker expression (alkaline phosphatase, Oct4, Sox2, SSEA-1).
- Culture and differentiation of germ cells isolated from embryoid bodies (EBs).
Main Results:
- Rat MPCs supported mouse Oct4-GFP ESC growth comparably to MEFs (MPC: 85.5% ± 5.1% vs. MEF: 84.1% ± 6.2%).
- ESCs maintained morphology and stem cell marker expression for over 15 passages on MPCs.
- Rat MPCs supported the survival and differentiation of germ cells into spermatogonial stem-like cells.
Conclusions:
- Rat MPCs provide an effective and simpler feeder system for culturing mouse ESCs and germ cells.
- This system offers a practical alternative to conventional MEF-based culture methods.
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