Isolation of adult pituitary stem/progenitor cell clusters located in the parenchyma of the rat anterior lobe.
Recent studies have demonstrated that Sox2-expressing stem/progenitor cells play roles in the pituitary cell turnover. Two types of niches have been proposed for stem/progenitor cells, the marginal cell layer (MCL) and the dense cell clusters in the parenchyma. Among them, the appearance of the parenchymal-niche only after birth indicates that this niche is involved in the cell turnover required for the postnatal pituitary. However, little is known about the roles of the parenchymal-niche and its regulation. The present study aimed to isolate pituitary stem/progenitor cells from the parenchymal-niche in the adult rat pituitary. Cell dispersion by stepwise treatment with proteases allowed the isolation of dense cell clusters. Immunocytochemistry demonstrated that clusters are universally composed of SOX2-positive cells, and most of them are positive for PROP1. Taken together with the anatomical analysis, we concluded that the isolated clusters are the parenchymal stem/progenitor cell (PS)-clusters, not the MCL-one. PS-clusters cultivated by serum-free overlay 3-dimensional culture maintained their stemness, and treatment with bFGF and EGF induced cyst-formation. Moreover, PS-clusters demonstrated some differentiation capacity with GSK3β-inhibitor treatment. Collectively, the present study demonstrates a simple method for isolating stem/progenitor cells from the parenchymal-niche, and provides tools to analyze the factors for regulating the pituitary niches.
Division of Life Science, Graduate School of Agriculture, Meiji University, Kanagawa, Japan; Organization for the Strategic Coordination of Research and Intellectual Property, Meiji University, Kanagawa, Japan; Institute of Reproduction and Endocrinology, Meiji University, Kanagawa, Japan.
Saishu
S
Yoshida
Division of Life Science, Graduate School of Agriculture, Meiji University, Kanagawa, Japan.
Naoto
N
Nishimura
Division of Life Science, Graduate School of Agriculture, Meiji University, Kanagawa, Japan.
Hiroki
H
Ueharu
Division of Life Science, Graduate School of Agriculture, Meiji University, Kanagawa, Japan.
Naoko
N
Kanno
Organization for the Strategic Coordination of Research and Intellectual Property, Meiji University, Kanagawa, Japan.
Masashi
M
Higuchi
Laboratory of Anatomy and Cell Biology, Department of Health Sciences, Kyorin University, Tokyo, Japan.
Kotaro
K
Horiguchi
Organization for the Strategic Coordination of Research and Intellectual Property, Meiji University, Kanagawa, Japan; Institute of Reproduction and Endocrinology, Meiji University, Kanagawa, Japan.
Takako
T
Kato
Division of Life Science, Graduate School of Agriculture, Meiji University, Kanagawa, Japan; Institute of Reproduction and Endocrinology, Meiji University, Kanagawa, Japan; Department of Life Science, School of Agriculture, Meiji University, Kanagawa, Japan. Electronic address: yukato@isc.meiji.ac.jp.
Yukio
Y
Kato
D053687:Adult Stem Cells / Q000166:cytology* / Q000378:metabolism; D000818:Animals; D015820:Cadherins / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D004815:Epidermal Growth Factor / Q000494:pharmacology; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D000071679:Glycogen Synthase Kinase 3 beta / Q000037:antagonists & inhibitors / Q000378:metabolism; D018398:Homeodomain Proteins / Q000378:metabolism; D007150:Immunohistochemistry; D008297:Male; D008856:Microscopy, Fluorescence; D010903:Pituitary Gland, Anterior / Q000166:cytology*; D051381:Rats; D055647:Rats, Transgenic; D017208:Rats, Wistar; D064568:S100 Calcium Binding Protein beta Subunit / Q000378:metabolism; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000378:metabolism
D015820:Cadherins; D018398:Homeodomain Proteins; C102134:Prophet of Pit-1 protein; D064568:S100 Calcium Binding Protein beta Subunit; D055748:SOXB1 Transcription Factors; D016222:Fibroblast Growth Factor 2; D004815:Epidermal Growth Factor; D000071679:Glycogen Synthase Kinase 3 beta
PROP1; Pituitary; S100β; SOX2-positive-cluster; Stem/progenitor cell niche