Differentiation of human mesenchymal stem cells into insulin-producing cells

• Main Authors: Ching-Fang Chang, 張靜芳
• Other Authors: Shih-Chieh Hung
• Format: Others
• Language: zh-TW
• Published: 2006
• Online Access: http://ndltd.ncl.edu.tw/handle/31686790954625708835

碩士 === 國立陽明大學 === 解剖暨細胞生物學研究所 === 95 === Type I diabetes mellitus is caused by an autoimmune destruction of the pancreatic islet β cells. The major obstacle in using trans- plantation for curing the disease is the limited source of insulin- producing cells. Stem cells represent a promising solution to this problem, and current research is being aimed at the creation of islet-endocrine tissue from those undifferentiated cells. Human mesenchymal stem cells(hMSCs)are self-renewing elements that can differentiated into multiple cell types ,including bone, cartilage, adipose and neuron. We present a method for forming insulin-producing cells derived from hMSCs. The protocol is consisted of several steps: Those hMSCs cultured in medium containing 3-Isobutyl-1-Methylxanthine (IBMX) and Insulin-Transferrin-Selenium-A(ITSA)would be induced to differentiate into neuron. Then the medium is replaced by fresh medium containing Nicotinamide, N2 and B27 would be induced to differentiate into pancreatic β cells. Our data shows that hMSCs cultured in medium containing extracellular matrix-fibronectin in suspension state would be differentiate into insulin-producing cells. Detecting insulin gene by Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR), we found those cells express insulin gene. Immuno- histochemistry stain shows those cells have high level proinsulin and insulin. In addition, ELISA data shows those cells secrete insulin amounted to 1.05 ±0.065ng/pellet/hr. However in animal model experiment, those cells can’t rescue the diabetic NOD/SCID mice. Future work will be the mechanism of extracellular matrix-fibronectin involved in stem cells differentiated into insulin-producing cells and establishing an effective animal model experiment.