Adult Mouse POMC/GFP
mHypoA-POMC/GFP-1-4 and mPitA-POMC/GFP-1
The mHypoA-POMC/GFP cell lines were generated using a proprietary platform technology enabling the creation of pools of immortalized hypothalamic neurons. These neurons were immortalized from four 8 week old male POMC-GFP mice (mouse 1-4) (strain C57BL/6J-Tg(Pomc-EGFP)1Low/J) hypothalamic primary cultures by retroviral transfer of SV40 T-Ag. These mice contain an eGFP transgene driven by the POMC promoter. The four immortalized hypothalamic pools were sorted on a cell sorting flow cytometer based on GFP fluorescence. Four cell lines representing the heterogeneous population of hypothalamic POMC neurons resulted: mHypoA-POMC/GFP lines 1–4.
These cells have been found to express an ever expanding array of neuropeptides, enzymatic markers and biologically active receptors. As such, these cells enable accurate in-vitro assays for use in the discovery, development and validation of new therapeutics targeted to central-nervous system diseases and disorders, including obesity, stress, and metabolic disorders, amongst others.
mPitA-POMC/GFP-1 was generated from a pool of immortalized pituitary neurons. These neurons were immortalized from 8 week old male POMC-GFP mouse (mouse 1) (strain C57BL/6J-Tg(Pomc-EGFP)1Low/J) pituitary primary cultures by retroviral transfer of SV40 T-Ag. The immortalized pituitary pool was sorted on a cell sorting flow cytometer based on GFP fluorescence as above. A cell line representing the heterogeneous population of POMC expressing pituitary neurons resulted: mPitA-POMC/GFP-1.
This pituitary cell line is easy to culture and has robust gene and protein expression. As such, this adult-derived pituitary cell line enables accurate assays for use in studies of progenitor cell characteristics and modulation, and the molecular analysis of hormone synthesis and secretion in differentiated cells.
Further details regarding these cell lines can be found in the following paper:
The datasheet for these products can be downloaded here.
The Immunocytochemistry (ICC) protocol for staining of ACTH/CLIP on mPitA-POMC/GFP-1 can be downloaded here.
Figure 1. The mHypoA-POMC/GFP cell lines co-express α-MSH and GFP. ICC was performed on each cell line with antibodies specific to α-MSH (red) and GFP (green). The nucleic acid stain TO-PRO-3 was used to visualize cell nuclei (blue). (Nazarians-Armavil, A. et al. (2014) Journal of Endocrinology 220(1): 13-24.)
Figure 2. The mPitA-POMC/GFP-1 cell line co-express ACTH/CLIP and GFP. ICC was performed on the cell line with antibodies specific to ACTH/CLIP (red) and GFP (green). The nucleic acid stain TO-PRO-3 was used to visualize cell nuclei (blue).
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