1.Effects of CXCR4 gene transfection on biologicalbehavior of bone marrow mesenchymal stem cells (in vitro)
Yuying WANG ; Nan ZHANG ; Xiuli LI ; Yameng WANG ; Shaoheng LI ; Yuhui YAN ; Jie SONG ; Jingxian YANG ; Qingping WEN
Chinese Pharmacological Bulletin 2017;33(6):806-813
Aim To observate the effect of chemokine receptor(CXCR4) gene transfection on biological behavior of bone marrow mesenchymal stem cells in vitro.Methods Firstly, bone marrow mesenchymal stem cells were divided into three groups:GFP(transfected GFP into MSCs), CXCR4+(transfected CXCR4+ into MSCs) and CXCR4-(transfected CXCR4-into MSCs) group.Then, their capacity of proliferation, differentiation and migration ability (in vitro) was assessed with immunofluorescence cytochemistry method, flow cytometry assay and Transwell cell chemotaxis test.Results The high or low expression of CXCR4 had no effect on their ability of proliferation and differentiation into lung tissue.Compared with GFP group, however, CXCR4+-MSCs group significantly increased the number of migrating cells, while CXCR4——MSCs group showed no change in the number of migrating cells.Conclusions The proliferation and differentiation capacities are not affected by the high or low expression of CXCR4.The high expression of CXCR4 can significantly enhance the migration ability of MSCs to inflammatory lesions, and the low one has no effect on the migration of the cells.After the transplantation of MSCs, CXCR4′s high expression will access to the lesion area to participate in tissue repairing rapidly and largely, significantly enhancing the therapeutic efficacy.
2.Clinical and pathological characteristics of 107 esophageal neuroendocrine carcinoma
Yu MENG ; Yanbang LIAN ; Shanshan LI ; He GOU ; Yameng WANG ; Jie CHEN ; Min SONG
Chinese Journal of Digestion 2019;39(8):533-538
Objective To investigate the clinical and pathological features of patients with esophageal neuroendocrine carcinoma (ENEC).Methods From January 2011 to November 2018,107 patients with pathologically confirmed ENEC were enrolled at the First Affiliated Hospital of Zhengzhou University.The clinical manifestation,tumor location,tumor size,clinical pathological classification and immunohistochemical markers were analyzed.Statistical description was used for measurement data analysis,and chi-square test was performed for classification data analysis.Results Among 107 patients with ENEC,feeling obstruction during eating was the most common initial symptom,accounting for 63.6% (68/107);followed by chest and back pain,accounting for 13.1% (14/107).About 60.7% (65/107) patients were diagnosed by biopsy under endoscopy and 39.3% (42/107) patients were confirmed by pathological diagnosis after surgery.The proportion of tumor located in the upper thoracic esophagus and middle and lower thoracic segments was 13.1% (14/107),45.8% (49/107) and 41.1% (44/107),respectively.The length of tumor was 0.7 cm to 9.0 cm,and the median was 2.5 cm.Among them,57.0% (61/107) were less than 2.5 cm and 43.0% (46/107) were over 2.5 cm.Among 107 patients,50 (46.7%) patients were ulcerative type,32 (29.9%) patients were medullary type,16 (15.0%) patients were mushroom type and nine (8.4%) patients were protrude type.Among 107 patients,96 (89.7%) patients were pure neuroendocrine carcinoma (P-NEC;including 95 small cell types,one large cell type);11 (10.3%) patients were mixed neuroendocrine carcinoma (M-NEC;including nine small cell carcinoma mixed with squamous cell carcinoma,two small cell carcinoma mixed with adenocarcinoma).The positive rates of synaptophysin,CD56 and chromogranin A were 99.0% (104/105),98.0% (100/102) and 31.5% (17/54),respectively.Ki-67 proliferation index of 47.7% tumors (51/107) was between 90% and 100%.P-NEC with the maximum diameter over 2.5 cm accounting for 42.1% (45/107),and M-NEC accounting for 0.9% (1/107).The maximum diameter of P-NEC group was larger than that of M-NEC group,and the difference was statistically significant (x2 =4.311,P =0.038).Conclusions ENEC is a kind of highly aggressive malignant tumor with nonspecific manifestations.The diagnosis mainly depends on histopathology and immunohistochemistry.
3.Define of Optimal Addition Period of Osteogenic Peptide to Accelerate the Osteogenic Differentiation of Human Pluripotent Stem Cells
Yameng SONG ; Hongjiao LI ; Zixuan WANG ; Jiamin SHI ; Jing LI ; Lu WANG ; Lingzi LIAO ; Shengqin MA ; Yun ZHANG ; Bin LIU ; Yaling YANG ; Ping ZHOU
Tissue Engineering and Regenerative Medicine 2024;21(2):291-308
BACKGROUND:
The addition of growth factiors is commonly applied to improve the osteogenic differentiation of stem cells. However, for human pluripotent stem cells (hPSCs), their complex differentiation processes result in the unknown effect at different stages. In this study, we focused on the widely used bone forming peptide-1 (BFP-1) and investigated the effect and mechanisms of its addition on the osteogenic induction of hPSCs as a function of the supplementation period.
METHODS:
Monolayer-cultured hPSCs were cultured in osteogenic induction medium for 28 days, and the effect of BFP-1 peptide addition at varying weeks was examined. After differentiation for varying days (0, 7, 14, 21 and 28), the differentiation efficiency was determined by RT–PCR, flow cytometry, immunofluorescence, and alizarin red staining assays. Moreover, the expression of marker genes related to germ layers and epithelial-mesenchymal transition (EMT) was investigated at day 7.
RESULTS:
Peptide treatment during the first week promoted the generation of mesoderm cells and mesenchymal-like cells from hiPSCs. Then, the upregulated expression of osteogenesis marker genes/proteins was detected in both hESCs and hiPSCs during subsequent inductions with BFP-1 peptide treatment. Fortunately, further experimental design confirmed that treating the BFP-1 peptide during 7–21 days showed even better performance for hESCs but was ineffective for hiPSCs.
CONCLUSION
The differentiation efficiency of cells could be improved by determining the optimal treatment period.Our study has great value in maximizing the differentiation of hPSCs by adding osteogenesis peptides based on the revealed mechanisms and promoting the application of hPSCs in bone tissue regeneration.
4.Advances in the differentiation of human induced pluripotent stem cells into osteoblasts
LIAO Lingzi ; SONG Yameng ; LIU Meixuan ; LI Siyi ; ZHOU Ping
Journal of Prevention and Treatment for Stomatological Diseases 2024;32(10):805-813
Bone diseases, such as osteoporosis and osteoarthritis, have emerged as pressing public health concerns requiring immediate attention and resolution. Cellular therapy and tissue engineering techniques are among the most promising therapeutic approaches for such conditions. Human induced pluripotent stem cells (hiPSCs) possess remarkable capacity for indefinite self-renewal in vitro and the ability to differentiate into all somatic cell types originating from the three germ layers, thereby making them a promising source of osteoblasts. Consequently, it is crucial to establish a well-delineated system for osteogenic differentiation of hiPSCs in vitro, with the aim to generate osteoblast-like cells that conform to clinical application standards. Numerous research teams have achieved substantial advancements in both the direct osteogenic differentiation of hiPSCs and the indirect pathway via mesenchymal stem cells. In this article, we provide a comprehensive review of these two osteogenic differentiation pathways and their current applications, with the aim of serving as a valuable reference for bone regeneration technologies. Current research efforts have relied on embryoid body formation and monolayer induction methods utilizing biomaterials to develop a system that facilitates in vitro culture and osteogenic differentiation of hiPSCs. However, the existing research is primarily constrained by unclear system components and low efficiency. Therefore, the development of a stepwise and three-dimensional induction system based on stringent regulation by specific compounds is a primary research direction for the future.