Isolation and characterization of stem cells from the human parathyroid gland.
OBJECTIVE
Somatic stem cells can be obtained from a variety of adult human tissues. However, it was not clear whether human parathyroid glands, which secrete parathyroid hormones and are essential in maintaining homeostasis levels of calcium ions in the circulation, contained stem cells. We aimed to investigate the possibility of isolating such parathyroid-derived stem cells (PDSC).


METHODS
Surgically removed parathyroid glands were obtained with informed consent. Cell cytogenetics was used to observe chromosomal abnormalities. Surface phenotypes were characterized by flow cytometry. Telomerase repeat amplification protocol (TRAP) assay was performed to observe the telomerase activity. RT-PCR and real-time PCR was was used to detect gene expressions. Real-time calcium uptake imaging was performed for extent of calcium uptake and transmission electron microscopy and immunofluorecent staining for smooth muscle actin.


RESULTS
After enzymatic digestion and primary culture, plastic-adherent, fibroblast-like cells appeared in culture and a morphologically homogeneous population was derived from subsequent limiting dilution and clonal expansion. Karyotyping was normal and doubling time of clonal cell growth was estimated to be 70.7 +/- 14.5 h (mean +/- standard deviation). The surface phenotype of the cells was positive for CD73, CD166, CD29, CD49a, CD49b, CD49d, CD44, CD105, and MHC class I, and negative for CD34, CD133, CD117, CD114, CD31, CD62P, EGF-R, ICAM-3, CD26, CXCR4, CD106, CD90 and MHC class II, similar to mesenchymal stem cells (MSC). Detectable levels of telomerase activity along with pluripotency Sall4 gene expression were observed from the isolated PDSCs. Expression of calcium-sensing receptor gene along with alpha-smooth muscle actin was induced and cellular uptake of extracellular calcium ions was observed. Furthermore, PDSCs possessed osteogenic, chondrogenic and adipogenic differentiation potentials.


CONCLUSIONS
Our results reveal that PDSCs were similar phenotypically to MSCs and further studies are needed to formulate induction conditions to differentiate PDSCs into parathyroid hormone-secreting chief cells.
Shih
Y-R V
YR
Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan.
Kuo
T K
TK
Yang
A-H
AH
Lee
O K
OK
Lee
C-H
CH
D000199:Actins / Q000032:analysis; D050156:Adipogenesis; D002118:Calcium / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002463:Cell Membrane Permeability; D049109:Cell Proliferation; D002469:Cell Separation; D002478:Cells, Cultured; D020219:Chondrogenesis; D015870:Gene Expression; D006801:Humans; D010012:Osteogenesis; D010280:Parathyroid Glands / Q000166:cytology* / Q000601:surgery; D010641:Phenotype; D044169:Receptors, Calcium-Sensing / Q000235:genetics; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism*; D019098:Telomerase / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics
D000199:Actins; C095550:CASR protein, human; D044169:Receptors, Calcium-Sensing; C472575:SALL4 protein, human; D014157:Transcription Factors; D019098:Telomerase; D002118:Calcium