A Novel Approach for Amplification and Purification of Mouse Oligodendrocyte Progenitor Cells

Yang, Junlin; Cheng, Xiaowen; Shen, Jiaxi; Xie, Binghua; Zhao, Xiaofeng; Zhang, Zunyi; Cao, Qi Lin; Shen, Ying; Qiu, Mengsheng

doi:10.3389/fncel.2016.00203

Cited by 31 publications

(28 citation statements)

References 39 publications

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“…Recently, a new method to enrich OPCs in mixed cultures from mouse cortices has been developed. Using a cocktail of the growth factors Platelet Derived Growth Factor-AA (PDGFaa), basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) OPCs could be propagated ( Yang et al, 2016 ). Interestingly, in mouse the efficiency of the OPC isolation is strongly correlated with the age of the donor tissue: the number of adult OPCs reaches 12% of the cell number that could be isolated from neonates ( Pillai and Ross, 1986 ).…”

Section: Discussionmentioning

confidence: 99%

Primary Spinal OPC Culture System from Adult Zebrafish to Study Oligodendrocyte Differentiation In Vitro

Kroehne

Tsata

Marrone

et al. 2017

Front. Cell. Neurosci.

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Endogenous oligodendrocyte progenitor cells (OPCs) are a promising target to improve functional recovery after spinal cord injury (SCI) by remyelinating denuded, and therefore vulnerable, axons. Demyelination is the result of a primary insult and secondary injury, leading to conduction blocks and long-term degeneration of the axons, which subsequently can lead to the loss of their neurons. In response to SCI, dormant OPCs can be activated and subsequently start to proliferate and differentiate into mature myelinating oligodendrocytes (OLs). Therefore, researchers strive to control OPC responses, and utilize small molecule screening approaches in order to identify mechanisms of OPC activation, proliferation, migration and differentiation. In zebrafish, OPCs remyelinate axons of the optic tract after lysophosphatidylcholine (LPC)-induced demyelination back to full thickness myelin sheaths. In contrast to zebrafish, mammalian OPCs are highly vulnerable to excitotoxic stress, a cause of secondary injury, and remyelination remains insufficient. Generally, injury induced remyelination leads to shorter internodes and thinner myelin sheaths in mammals. In this study, we show that myelin sheaths are lost early after a complete spinal transection injury, but are re-established within 14 days after lesion. We introduce a novel, easy-to-use, inexpensive and highly reproducible OPC culture system based on dormant spinal OPCs from adult zebrafish that enables in vitro analysis. Zebrafish OPCs are robust, can easily be purified with high viability and taken into cell culture. This method enables to examine why zebrafish OPCs remyelinate better than their mammalian counterparts, identify cell intrinsic responses, which could lead to pro-proliferating or pro-differentiating strategies, and to test small molecule approaches. In this methodology paper, we show efficient isolation of OPCs from adult zebrafish spinal cord and describe culture conditions that enable analysis up to 10 days in vitro. Finally, we demonstrate that zebrafish OPCs differentiate into Myelin Basic Protein (MBP)-expressing OLs when co-cultured with human motor neurons differentiated from induced pluripotent stem cells (iPSCs). This shows that the basic mechanisms of oligodendrocyte differentiation are conserved across species and that understanding the regulation of zebrafish OPCs can contribute to the development of new treatments to human diseases.

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Section: Discussionmentioning

confidence: 99%

Primary Spinal OPC Culture System from Adult Zebrafish to Study Oligodendrocyte Differentiation In Vitro

Kroehne

Tsata

Marrone

et al. 2017

Front. Cell. Neurosci.

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“…Tripotential glial-restricted precursor (GRP) cells were initially found in embryonic spinal tissues (Herrera et al, 2001; Gregori et al, 2002; Wu et al, 2002; Cao et al, 2005; Phillips et al, 2012), as they can generate OLs and type I and type II astrocytes in vitro (Rao et al, 1998; Gregori et al, 2002; Yang et al, 2016) and in vivo (Herrera et al, 2001; Hill et al, 2004). The germination of GRP cells from neuroepithelial stem cells was viewed as the beginning of OL generation (Rao et al, 1998; Gregori et al, 2002).…”

Section: Introductionmentioning

confidence: 99%

EGF Enhances Oligodendrogenesis from Glial Progenitor Cells

Yang

Cheng

et al. 2017

Front. Mol. Neurosci.

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Emerging evidence indicates that epidermal growth factor (EGF) signaling plays a positive role in myelin development and repair, but little is known about its biological effects on the early generation and differentiation of oligodendrocyte (OL) lineage cells. In this study, we investigated the role of EGF in early OL development with isolated glial restricted precursor (GRP) cells. It was found that EGF collaborated with Platelet Derived Growth Factor-AA (PDGFaa) to promote the survival and self-renewal of GRP cells, but predisposed GRP cells to develop into O4− early-stage oligodendrocyte precursor cells (OPCs) in the absence of or PDGFaa. In OPCs, EGF synergized with PDGFaa to maintain their O4 negative antigenic phenotype. Upon PDGFaa withdrawal, EGF promoted the terminal differentiation of OPCs by reducing apoptosis and increasing the number of mature OLs. Together, these data revealed that EGF is an important mitogen to enhance oligodendroglial development.

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“…Several strategies for purifying OPCs have been implemented, such as immunopanning [10], magnetic activated cell sorting [11], fluorescence-activated cell sorting [12], special factors induced approach [13], differential and density gradient centrifugation, and shaking method.…”

Section: Effects Of Different Purified Strategies On Isolation Of Opcsmentioning

confidence: 99%

Reprogramming of mouse embryonic fibroblasts into induced oligodendrocyte progenitor cells via nanovector-mediated indirect lineage conversion

Xiao

Ling

Qiu

et al. 2020

Preprint

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Background: Axon-wrapping myelin sheath is vital to nerve conduction, loss or dysfunction of it will result in some neurological diseases. Since mature oligodendrocytes (OLs) lack proliferation ability, and oligodendrocyte progenitor cells (OPCs) maintain slow proliferation or quiescence state, transplantation of exogenous OPCs becomes an alternative strategy for curing these diseases. Methods: OPCs were isolated from brain tissue and generated from NIH/3T3 cells via indirect lineage conversion here. Results: 0.25 % chicken serum was conducive to trypsin-mediated digestion of cerebral cortex, the modified shaking method could improve the purification efficiency of OPCs. Poly-MAG was a safer and higher efficient nanovector for delivering plasmid DNA into cells. Forced expressions of Olig 2, Nkx 6.2, and Sox 10 could attenuate the expressions of fibroblast-related genes (Col5a1 and Col1a1), up-regulate OPC-associated genes (PDGFRα, S100β, NG2, and Olig 2), and finally successfully reprogram NIH/3T3 cells into induced oligodendrocyte progenitor cells (iOPCs). 1 μM haloperidol (HAL) contributed to promoting the proliferation of iOPCs. These iOPCs had the potential to be differentiated into OLs and type Ⅱ astrocytes. Conclusions: The high-efficiency and low-cytotoxicity strategies for isolating mouse primary OPCs and generating iOPCs were established, which provided novel cell resources for disease modeling, drug screening, cell therapy, and so on.

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A Novel Approach for Amplification and Purification of Mouse Oligodendrocyte Progenitor Cells

Cited by 31 publications

References 39 publications

Primary Spinal OPC Culture System from Adult Zebrafish to Study Oligodendrocyte Differentiation In Vitro

Primary Spinal OPC Culture System from Adult Zebrafish to Study Oligodendrocyte Differentiation In Vitro

EGF Enhances Oligodendrogenesis from Glial Progenitor Cells

Reprogramming of mouse embryonic fibroblasts into induced oligodendrocyte progenitor cells via nanovector-mediated indirect lineage conversion

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