The Establishment of In Vitro Human Induced Pluripotent Stem Cell-Derived

被引:0
作者
Idris, Izyan Mohd [1 ,2 ]
Nordin, Fazlina [1 ]
Muhamad, Nur Jannaim [2 ]
Jalil, Julaina Abdul [2 ]
Diana, Fatimah
Ordin, Aminn [2 ]
Mohamed, Rosnani [2 ]
Matripen, Adiratna [2 ]
Tye, Gee Jun
Zaman, Wan Safwani Wankamarul [3 ,4 ]
Yazid, Muhammad Dain
Hweing, Min [1 ]
机构
[1] Univ Kebangsaan Malaysia Med Ctr UKMMC, Ctr Tissue Engn & Regenerat Med CTERM, Fed Terr, Kuala Lumpur 56000, Malaysia
[2] Minist Hlth Malaysia, Inst Med Res IMR, Natl Inst Hlth NIH, Jalan Setia Murni U13-52, Seksyen U13 Setia Alam, Shah Alam 40170, Selangor, Malaysia
[3] Univ Sains Malaysia USM, Inst Res Mol Med INFORMM, USM, George Town 11800, Malaysia
[4] Univ Malaya, Fac Engn, Dept Biomed Engn, Fed Terr, Kuala Lumpur 50603, Malaysia
来源
SAINS MALAYSIANA | 2023年 / 52卷 / 03期
关键词
Induced pluripotent stem cells; lentivirus; neural stem cells; neurons; reprogramming; HUMAN FIBROBLASTS; GENERATION; EFFICIENCY; INDUCTION; CULTURE; VECTOR; GROWTH;
D O I
10.17576/jsm-2023-5203-14
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Induced pluripotent stem cells (iPSCs) have been generated using different reprogramming strategies. Lentiviruses remain a strategic method for cell reprogramming as it is highly efficient in gene transfer. The latest fourth-generation lentiviral packaging systems claimed to be efficient and safe. However, modifications made to enhance safety of lentiviral vectors have been shown to affect vector performance. In this study, we established that the fourthgeneration lentiviral packaging system can produce high-titre lentiviruses with high-transduction efficiencies. Subsequently, the robustness and reproducibility of generating iPSCs from adult human dermal fibroblasts were tested using these lentiviruses. The use of fourth-generation lentiviruses consistently generates iPSCs with similar efficiency and quality in different primary cell lines. This study demonstrated that the human-derived iPSCs can be maintained using mitomycin-C inactivated feeder cells. The iPSCs clones highly expressed key pluripotency markers and can spontaneously differentiate into cells from the three embryonic germ layers. The iPSCs generated were able to differentiate into neural stem cell lineages, producing cells expressing Nestin and Sox2 as well as able to further differentiate into neurons with more than 70% efficiency. The data demonstrated that the use of the fourth-generation lentiviral packaging to produce lentiviruses for iPSCs generation is robust and reproducible as it can generate iPSCs from different adult dermal fibroblasts with the potential to differentiate into neural stem cells and neurons. The use of safer lentiviral packaging systems combined with established vector plasmids will help to expedite the generation of iPSCs for clinical applications.
引用
收藏
页码:863 / 876
页数:14
相关论文
共 36 条
[1]   Efficient and rapid generation of induced pluripotent stem cells from human keratinocytes [J].
Aasen, Trond ;
Raya, Angel ;
Barrero, Maria J. ;
Garreta, Elena ;
Consiglio, Antonella ;
Gonzalez, Federico ;
Vassena, Rita ;
Bilic, Josipa ;
Pekarik, Vladimir ;
Tiscornia, Gustavo ;
Edel, Michael ;
Boue, Stephanie ;
Izpisua Belmonte, Juan Carlos .
NATURE BIOTECHNOLOGY, 2008, 26 (11) :1276-1284
[2]  
Andrews PW, 2015, REGEN MED, V10, P1, DOI [10.2217/RME.14.93, 10.2217/rme.14.93]
[3]   Impact of heterogeneous perisomatic IPSC populations on pyramidal cell firing rates [J].
Aradi, I ;
Santhakumar, V ;
Soltesz, I .
JOURNAL OF NEUROPHYSIOLOGY, 2004, 91 (06) :2849-2858
[4]   Multipotent cell lineages in early mouse development depend on SOX2 function [J].
Avilion, AA ;
Nicolis, SK ;
Pevny, LH ;
Perez, L ;
Vivian, N ;
Lovell-Badge, R .
GENES & DEVELOPMENT, 2003, 17 (01) :126-140
[5]   Differentiation of Human Induced Pluripotent Stem Cells (iPSCs) into an Effective Model of Forebrain Neural Progenitor Cells and Mature Neurons [J].
Bell, Scott ;
Hettige, Nuwan C. ;
Silveira, Heika ;
Peng, Huashan ;
Wu, Hanrong ;
Jefri, Malvin ;
Antonyan, Lilit ;
Zhang, Ying ;
Zhang, Xin ;
Ernst, Carl .
BIO-PROTOCOL, 2019, 9 (05)
[6]   Progress and Perspectives in the Development of Lentiviral Vector Producer Cells [J].
Ferreira, Mariana, V ;
Cabral, Elisa T. ;
Coroadinha, Ana Sofia .
BIOTECHNOLOGY JOURNAL, 2021, 16 (01)
[7]   Efficient induction of transgene-free human pluripotent stem cells using a vector based on Sendai virus, an RNA virus that does not integrate into the host genome [J].
Fusaki, Noemi ;
Ban, Hiroshi ;
Nishiyama, Akiyo ;
Saeki, Koichi ;
Hasegawa, Mamoru .
PROCEEDINGS OF THE JAPAN ACADEMY SERIES B-PHYSICAL AND BIOLOGICAL SCIENCES, 2009, 85 (08) :348-362
[8]   Methods for making induced pluripotent stem cells: reprogramming a la carte [J].
Gonzalez, Federico ;
Boue, Stephanie ;
Izpisua Belmonte, Juan Carlos .
NATURE REVIEWS GENETICS, 2011, 12 (04) :231-242
[9]   Pluripotent stem cells induced from adult neural stem cells by reprogramming with two factors [J].
Kim, Jeong Beom ;
Zaehres, Holm ;
Wu, Guangming ;
Gentile, Luca ;
Ko, Kinarm ;
Sebastiano, Vittorio ;
Arauzo-Bravo, Marcos J. ;
Ruau, David ;
Han, Dong Wook ;
Zenke, Martin ;
Schoeler, Hans R. .
NATURE, 2008, 454 (7204) :646-U54
[10]   Transgene-Free Disease-Specific Induced Pluripotent Stem Cells from Patients with Type 1 and Type 2 Diabetes [J].
Kudva, Yogish C. ;
Ohmine, Seiga ;
Greder, Lucas V. ;
Dutton, James R. ;
Armstrong, Adam ;
De Lamo, Josep Genebriera ;
Khan, Yulia Krotova ;
Thatava, Tayaramma ;
Hasegawa, Mamoru ;
Fusaki, Noemi ;
Slack, Jonathan M. W. ;
Ikeda, Yasuhiro .
STEM CELLS TRANSLATIONAL MEDICINE, 2012, 1 (06) :451-461