1.Role of adipose-derived mesenchymal stem cells in bone tissue engineering
Jinghan JIA ; Zhi ZHANG ; Xiangpeng WANG ; Dongqing ZOU
Chinese Journal of Tissue Engineering Research 2010;14(1):166-170
BACKGROUND: At present, bone defects usually repaired by autologous bone, allogenic bone, synthetic bone substitutes and other methods, which received poor clinical results. Preliminary studies have shown that adipose-derived mesenchymal stem cells (ADSCs) possess strong proliferation ability and differentiation potential, and can be induced differentiate into bone. OBJECTIVE: To analyze the application of ADSCs in bone tissue engineering, and to identify whether ADSCs can be used as seed cells in bone tissue engineering. METHODS: The databases of PubMed (1999-01/2008-12) and Tongfang (2003-01/2008-12) was retrieved using key words of "adipose tissue-derived mesenchymal stem cells, adipose mesenchymal stem cells, adipose stem cell; osteogenic induction, osteogenic inducement, bone induction, osteoblastic induced; chondroblast induction, cartilage induction; bone tissue engineering, tissue engineering bone, tissue engineering of bone". RESULTS AND CONCLUSION: A total of 361 literatures were collected, including 246 in Chinese and 115 in English. Totally 29 literatures were accordant with the study criteria. ADSCs is a truly multi-directional differentiation potential cells, which possess strong amplification and self-renewal potential, and can be directional differentiated into osteoblasts, cartilage cells, bone cells and muscle cells. It can be used as seed cells in bone tissue engineering when matching appropriate stents.
2.Generation of an induced pluripotent stem cell line from a patient with surfactant metabolism dysfunction carrying ABCA3 mutations
Zhichen TIAN ; Xin XIE ; Jinghan CHI ; Jia CHEN ; Danhua ZHAO ; Yanmei HE ; Xiaojuan YIN
Chinese Journal of Applied Clinical Pediatrics 2024;39(2):98-103
Objective:Induced pluripotent stem cells (iPSCs) cell lines were established using peripheral blood mononuclear cells (PBMCs) from a patient suffering from neonatal respiratory distress syndrome (NRDS) who carried Adenosine triphosphate-binding cassette transporter A3 ( ABCA3) compound heterozygous mutations. Methods:Cell experimental research.Peripheral venous blood was collected and PBMCs were isolated and cultured in vitro. PBMCs were transfected with non-integrated Sendai vector carrying reprogramming factors.The chromosome karyotypes of the established iPSCs were analyzed.Immunofluorescence and flow cytometry were used to detect pluripotency markers of stem cells and verify their differentiation potential.Sanger sequencing was performed to analyze gene mutations.In addition, short tandem repeat (STR) analysis was performed, polymerase chain reaction(PCR) and agarose gel electrophoresis were used to detect virus residual. Results:Karyotype analysis of established iPSCs cell lines showed normal diploid 46, XY karyotype.Immunofluorescence showed positive staining of stem cell pluripotency markers OCT4, SSEA4, Nanog and Sox2.Flow cytometry was used to detected stem cell pluripotency markers and showed expression of TRA-1-60, SSEA-4 and OCT4.After differentiation into all three germ layers, immunofluorescence was performed to detect ectoderm (Pax-6), mesoderm (Brachyury) and endoderm alpha-fetoprotein markers, and the results showed positive staining, which confirmed that the iPSCs had the potential to differentiate.Sanger sequencing showed c. 3997_3998del and c. 3137C>T compound heterozygous mutations.STR analysis showed they originate from PBMCs, and no Sendai virus residual was detected by PCR and agarose gel electrophoresis.Conclusions:In this study, PBMCs from patient carrying ABCA3 compound heterozygous mutations was used to establish iPSCs cell lines.The research lays a foundation for the study of pathogenesis, therapeutic drug screening and cell therapy of NRDS caused by ABCA3 gene mutations.