Chinese Journal of Tissue Engineering Research ›› 2024, Vol. 28 ›› Issue (25): 3942-3946.doi: 10.12307/2024.178

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Culture, identification, and differentiation of rabbit pituitary stem cells

Yue Tingting1, Zhu Ting2, Mao Jiannan1, Li Wei1, Hang Chunhua1, 2   

  1. 1Department of Neurosurgery, Nanjing Drum Tower Hospital Affiliated to Nanjing University Medical School, Nanjing 210008, Jiangsu Province, China; 2Nanjing Drum Tower Hospital Clinical College of Nanjing University of Chinese Medicine, Nanjing 210008, Jiangsu Province, China
  • Received:2023-05-22 Accepted:2023-07-04 Online:2024-09-08 Published:2023-11-23
  • Contact: Hang Chunhua, MD, Chief physician, Professor, Department of Neurosurgery, Nanjing Drum Tower Hospital Affiliated to Nanjing University Medical School, Nanjing 210008, Jiangsu Province, China; Nanjing Drum Tower Hospital Clinical College of Nanjing University of Chinese Medicine, Nanjing 210008, Jiangsu Province, China
  • About author:Yue Tingting, MD, Department of Neurosurgery, Nanjing Drum Tower Hospital Affiliated to Nanjing University Medical School, Nanjing 210008, Jiangsu Province, China
  • Supported by:
    National Natural Science Foundation of China (General Program), No. 81971122 (to HCH)

Abstract: BACKGROUND: The pituitary gland is an important endocrine organ in the body. Certain diseases can cause damage to the pituitary gland, such as pituitary adenoma and abnormal hormone secretion. Pituitary stem cells, due to their self-renewal and multi-directional differentiation potential, are expected to become a new therapeutic approach for repairing damaged pituitary glands.

OBJECTIVE: To isolate and culture pituitary stem cells using the suspension cell ball culture method and identify their proliferation and differentiation ability. 

METHODS: Pituitary stem cells were isolated and cultured from the pituitary gland of newborn New Zealand white rabbits using the suspension cell ball culture method, and their morphological characteristics were observed. Immunofluorescence cytochemistry was used to detect the expression of pituitary stem cell markers SOX2 and Nestin. EdU labeling method was utilized to detect the proliferative ability of pituitary stem cells. After adherent and induced differentiation, the hormone levels in the culture medium were detected by ELISA.

RESULTS AND CONCLUSION: Pituitary stem cell spheres could be successfully isolated by the suspension cell ball culture method, with strong proliferative ability. Positive expression of stem cell-specific markers SOX2 and Nestin was found in the cultured cells. After induction and differentiation, adrenocorticotropic hormone, thyroid hormone, growth hormone, luteinizing hormone, follicle-stimulating hormone, and prolactin levels significantly increased in the medium (P < 0.001), with strong differentiation ability.

Key words: pituitary stem cell, pituitary, suspension cell ball culture, appraisal, differentiation, rabbit

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