Developments in cell culture systems for human pluripotent stem cells

Liu, Weiwei; Deng, Chunhao; Godoy-Parejo, Carlos; Zhang, Yumeng; Chen, Guokai

doi:10.4252/wjsc.v11.i11.968

Cited by 15 publications

(11 citation statements)

References 124 publications

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“…Pluripotent stem cells (PSCs) can self-renew and differentiate to all cell types in the body. Specific mitogenic stimulations lead to specific pluripotency states such as naive and primed states that correspond to preimplant inner cell mass and postimplant epiblast during embryogenesis, respectively (Liu et al, 2019;Ying and Smith, 2017). Fibroblast growth factor 2 (FGF2) and transforming growth factor b (TGF-b) activation are required to maintain the primed state of human PSCs (hPSCs), including human embryonic stem cells (hESCs) (Thomson et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

“…Those serum products provide large amounts of uncharacterized lipids. With better understanding of pluripotency regulation, feeder-free culture systems have been developed to maintain hESCs with defined growth factors and different lipid supplements (Liu et al, 2019). Most feeder-free conditions contain chemically defined lipid concentrate (CDL), a synthetic product that contains eight common free fatty acids, cholesterol, and an antioxidant tocopherol (Figure S1A).…”

Section: Introductionmentioning

confidence: 99%

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Lysophosphatidic acid shifts metabolic and transcriptional landscapes to induce a distinct cellular state in human pluripotent stem cells

Xu¹,

Deng²,

Ren³

et al. 2021

Cell Reports

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Pluripotent stem cells (PSCs) can be maintained in a continuum of cellular states with distinct features. Exogenous lipid supplements can relieve the dependence on de novo lipogenesis and shift global metabolism. However, it is largely unexplored how specific lipid components regulate metabolism and subsequently the pluripotency state. In this study, we report that the metabolic landscape of human PSCs (hPSCs) is shifted by signaling lipid lysophosphatidic acid (LPA), which naturally exists. LPA leads to a distinctive transcriptome profile that is not associated with de novo lipogenesis. Although exogenous lipids such as cholesterol, common free fatty acids, and LPA can affect cellular metabolism, they are not necessary for maintaining primed pluripotency. Instead, LPA induces distinct and reversible phenotypes in cell cycle, morphology, and mitochondria. This study reveals a distinct primed state that could be used to alter cell physiology in hPSCs for basic research and stem cell applications.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Lysophosphatidic acid shifts metabolic and transcriptional landscapes to induce a distinct cellular state in human pluripotent stem cells

Xu¹,

Deng²,

Ren³

et al. 2021

Cell Reports

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“…Therefore, the morphology of hiPSC colonies is considered to be an important factor that indicates the pluripotency hiPSCs in vitro . HiPSCs cultured in vitro are usually passaged as multi-cellular clusters since single cells are more prone to cell death whereas colonies are quickly re-established by cell clusters [ 46 ]. We investigated the morphology of hiPSCs cultivated on MG + and MG − TA DMA and TB DMAs ( Fig.…”

Section: Resultsmentioning

confidence: 99%

Miniaturized droplet microarray platform enables maintenance of human induced pluripotent stem cell pluripotency

Liu

Chakraborty

Direksilp

et al. 2021

Materials Today Bio

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The capacity of human induced pluripotent stem cells (hiPSCs) for indefinite self-renewal warrants their application in disease modeling, drug discovery, toxicity assays and efficacy screening. However, their poor proliferation ability, inability to adhere to surfaces without Matrigel coating and tendency to spontaneously differentiate in vitro hinder the application of hiPSCs in these fields. Here we study the ability to culture hiPSCs inside 200 nL droplets on the droplet microarray (DMA) platform. We demonstrate that (1) hiPSCs can attach to the Matrigel (MG)-free surface of DMA and show good viability after 24 h culture; (2) hiPSC do not spontaneously differentiate when cultured on the MG-free surface of DMAs; (3) culturing of hiPSCs in 200 nL as compared to 2 mL culture leads to higher expression of the Nanog pluripotency marker. Overall, the results demonstrate the possibility to culture undifferentiated hiPSCs in 200 nL droplets on DMA, thereby opening the possibility for high-throughput screenings of hiPSCs with various factors without compromising the results through the involvement of animal-derived materials, such as Matrigel.

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“…Over the past two decades, culture conditions have been a major focus for hPSC research [3,130]. Recently, extended or expanded pluripotent stem cells (EPSCs) have been reported to have the additional ability to contribute to both embryonic and extraembryonic tissues [131][132][133].…”

Section: Hpscsmentioning

confidence: 99%

Applications of piggyBac Transposons for Genome Manipulation in Stem Cells

Liu

Huang

2021

Stem Cells International

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Transposons are mobile genetic elements in the genome. The piggyBac (PB) transposon system is increasingly being used for stem cell research due to its high transposition efficiency and seamless excision capacity. Over the past few decades, forward genetic screens based on PB transposons have been successfully established to identify genes associated with drug resistance and stem cell-related characteristics. Moreover, PB transposon is regarded as a promising gene therapy vector and has been used in some clinically relevant stem cells. Here, we review the recent progress on the basic biology of PB, highlight its applications in current stem cell research, and discuss its advantages and challenges.

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Developments in cell culture systems for human pluripotent stem cells

Cited by 15 publications

References 124 publications

Lysophosphatidic acid shifts metabolic and transcriptional landscapes to induce a distinct cellular state in human pluripotent stem cells

Lysophosphatidic acid shifts metabolic and transcriptional landscapes to induce a distinct cellular state in human pluripotent stem cells

Miniaturized droplet microarray platform enables maintenance of human induced pluripotent stem cell pluripotency

Applications of piggyBac Transposons for Genome Manipulation in Stem Cells

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