Proportion and morphological characteristics of human oligodendrocyte precursor cells in different cell culture vessels

doi:10.3969/j.issn.2095-4344.2139

Abstract

Abstract: Abstract
BACKGROUND: Oligodendrocyte precursor cell transplantation is one of the keys to the treatment of white matter damage in premature infants. At present, there is a lack of research on the comparison of oligodendrocyte precursor cells induced by neural stem cells derived from human fetal brain cultured in vitro in different vessels worldwide.
OBJECTIVE: To observe the morphology of human oligodendrocyte progenitor cells and pre-oligodendrocytes in different cell culture vessels (6-well plates, 24-well plates and T25 flasks).
METHODS: The 6-well plates, 24-well plates and T25 flasks were used as culture vessels to culture human oligodendrocyte progenitor cells and pre-oligodendrocytes. The characteristics of human oligodendrocyte progenitor cells were identified by immunofluorescence staining and flow cytometry. The morphology of cells was observed by an ordinary light microscope. Cell counts were performed according to cell morphology and statistical analysis was performed.
RESULTS AND CONCLUSION: (1) The oligodendrocyte progenitor cell body was round and the bipolar protrusions were bead-like; the pre-oligodendrocyte protrusions were more than two poles, and did not bifurcate. (2) The ratios of oligodendrocyte progenitor cell morphology in the oligodontia lines were significantly higher in the 6-well plates than those in the 24-well plates and T25 flasks (P < 0.05), followed by T25 flasks and 24-well plates. Morphological ratios of pre-oligodendrocytes were significantly higher in the 24-well plates compared to the 6-well plates and T25 flasks (P < 0.01), followed by T25 flasks and 6-well plates. (3) The cells cultured in the 6-well plate had fewer dregs, and the morphology, vigor and growth were better than those of the other culture vessels. (4) According to morphological view, 6-well plates are more suitable for oligodendrocyte progenitor cell growth, and 24-well plates are more suitable for pre-oligodendrocytes growth.

Key words: stem cells">, , oligodendrocyte precursor cells">, , pre-oligodendrocytes">, , cell morphology">, , cell viability

CLC Number:

Ye Dou, , Ma Xuexia , Guan Qian, , Luan Zuo , Yang Yinxiang , Wang Zhaoyan , Wang Qian , He Ying , Yao Ruiqin. Proportion and morphological characteristics of human oligodendrocyte precursor cells in different cell culture vessels[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(1): 44-49.

Figures/Tables 7

References

[1] VAN TILBORG E, DE THEIJE CGM, VAN HAL M, et al. Origin and dynamics of oligodendrocytes in the developing brain: Implications for perinatal white matter injury. Glia. 2018;66(2): 221-238.
[2] BACK SA. White matter injury in the preterm infant: pathology and mechanisms. Acta Neuropathol. 2017;134(3):331-349.
[3] BARATEIRO A, BRITES D, FERNANDES A. Oligodendrocyte Development and Myelination in Neurodevelopment: Molecular Mechanisms in Health and Disease. Curr Pharm Des. 2016;22(6):656-679.
[4] SALMASO N, JABLONSKA B, SCAFIDI J, et al. Neurobiology of premature brain injury. Nat Neurosci. 2014;17(3):341-346.
[5] ALIZADEH A, DYCK SM, KARIMI-ABDOLREZAEE S. Myelin damage and repair in pathologic CNS: challenges and prospects. Front Mol Neurosci. 2015;8:35.
[6] DIETZ KC, POLANCO JJ, POL SU, et al. Targeting human oligodendrocyte progenitors for myelin repair. Exp Neurol. 2016;283(Pt B):489-500.
[7] WANG S, BATES J, LI X, et al. Human iPSC-derived oligodendrocyte progenitor cells can myelinate and rescue a mouse model of congenital hypomyelination. Cell Stem Cell. 2013;12(2):252-264.
[8] YAMASHITA T, MIYAMOTO Y, BANDO Y, et al. Differentiation of oligodendrocyte progenitor cells from dissociated monolayer and feeder-free cultured pluripotent stem cells. PLoS One. 2017;12(2): e0171947.
[9] MCLANE LE, BOURNE JN, EVANGELOU AV, et al. Loss of Tuberous Sclerosis Complex1 in Adult Oligodendrocyte Progenitor Cells Enhances Axon Remyelination and Increases Myelin Thickness after a Focal Demyelination. J Neurosci. 2017;37(31):7534-7546.
[10] WANG C, LUAN Z, YANG Y, et al. High purity of human oligodendrocyte progenitor cells obtained from neural stem cells: suitable for clinical application. J Neurosci Methods. 2015;240:61-66.
[11] BUNGE RP. Glial cells and the central myelin sheath. Physiol Rev. 1968; 48(1):197-251.
[12] FILLEY CM, FIELDS RD. White matter and cognition: making the connection. J Neurophysiol. 2016;116(5):2093-2104.
[13] LESCA G, VANIER MT, CREISSON E, et al. X-linked adrenoleukodystrophy in a female proband: clinical presentation, biological diagnosis and family consequences. Arch Pediatr. 2005;12(8):1237-1240.
[14] Van der Knaap MS, Bugiani M. Leukodystrophies: a proposed classification system based on pathological changes and pathogenetic mechanisms. Acta Neuropathol. 2017;134(3): 351-382.
[15] KEOUGH MB, YONG VW. Remyelination therapy for multiple sclerosis. Neurotherapeutics. 2013;10(1):44-54.
[16] NISHIYAMA A, KOMITOVA M, SUZUKI R, et al. Polydendrocytes (NG2 cells): multifunctional cells with lineage plasticity. Nat Rev Neurosci. 2009;10(1):9-22.
[17] PAYNE SC, BARTLETT CA, SAVIGNI DL, et al. Early proliferation does not prevent the loss of oligodendrocyte progenitor cells during the chronic phase of secondary degeneration in a CNS white matter tract. PLoS One. 2013;8(6):e65710.
[18] ESPINOSA-JEFFREY A, BLANCHI B, BIANCOTTI JC, et al. Efficient Generation of Viral and Integration-Free Human Induced Pluripotent Stem Cell-Derived Oligodendrocytes. Curr Protoc Stem Cell Biol. 2016; 39(1):2D.18.1-2D.18.28.
[19] HU BY, DU ZW, ZHANG SC. Differentiation of human oligodendrocytes from pluripotent stem cells. Nat Protoc. 2009;4(11):1614-1622.
[20] SANTOS AK, VIEIRA MS, VASCONCELLOS R, et al. Decoding cell signalling and regulation of oligodendrocyte differentiation. Semin Cell Dev Biol. 2019;95:54-73.
[21] NEWVILLE J, JANTZIE LL, CUNNINGHAM LA.Embracing oligodendrocyte diversity in the context of perinatal injury. Neural Regen Res. 2017; 12(10):1575-1585.
[22] BARATEIRO A, FERNANDES A. Temporal oligodendrocyte lineage progression: in vitro models of proliferation, differentiation and myelination. Biochim Biophys Acta. 2014;1843(9):1917-1929.
[23] HUGHES EG, KANG SH, FUKAYA M, et al. Oligodendrocyte progenitors balance growth with self-repulsion to achieve homeostasis in the adult brain. Nat Neurosci. 2013;16(6): 668-676.
[24] BERGLES DE, RICHARDSON WD. Oligodendrocyte Development and Plasticity. Cold Spring Harb Perspect Biol. 2015;8(2):a020453.
[25] YEUNG MS, ZDUNEK S, BERGMANN O, et al. Dynamics of oligodendrocyte generation and myelination in the human brain. Cell. 2014;159(4):766-774.
[26] EL WALY B, MACCHI M, CAYRE M, et al. Oligodendrogenesis in the normal and pathological central nervous system. Front Neurosci. 2014; 8:145.
[27] NAVE KA, WERNER HB. Myelination of the nervous system: mechanisms and functions. Annu Rev Cell Dev Biol. 2014;30: 503-533.
[28] BULLER B, CHOPP M, UENO Y, et al. Regulation of serum response factor by miRNA-200 and miRNA-9 modulates oligodendrocyte progenitor cell differentiation. Glia. 2012; 60(12):1906-1914.
[29] MCMURRAN CE, KODALI S, YOUNG A, et al. Clinical implications of myelin regeneration in the central nervous system. Expert Rev Neurother. 2018;18(2):111-123.
[30] CZEPIEL M, BODDEKE E, COPRAY S. Human oligodendrocytes in remyelination research. Glia. 2015;63(4): 513-530.
[31] OSORIO MJ, ROWITCH DH, TESAR P, et al. Concise Review: Stem Cell-Based Treatment of Pelizaeus- Merzbacher Disease. Stem Cells. 2017; 35(2):311-315.
[32] DOUVARAS P, RUSIELEWICZ T, KIM KH, et al. Epigenetic Modulation of Human Induced Pluripotent Stem Cell Differentiation to Oligodendrocytes. Int J Mol Sci. 2016;17(4). pii: E614.
[33] THIRUVALLUVAN A, CZEPIEL M, KAP YA, et al. Survival and Functionality of Human Induced Pluripotent Stem Cell-Derived Oligodendrocytes in a Nonhuman Primate Model for Multiple Sclerosis. Stem Cells Transl Med. 2016;5(11): 1550-1561.
[34] PIAO J, MAJOR T, AUYEUNG G, et al. Human embryonic stem cell-derived oligodendrocyte progenitors remyelinate the brain and rescue behavioral deficits following radiation. Cell Stem Cell. 2015;16(2): 198-210.
[35] MARTEYN A, SARRAZIN N, YAN J, et al. Modulation of the Innate Immune Response by Human Neural Precursors Prevails over Oligodendrocyte Progenitor Remyelination to Rescue a Severe Model of Pelizaeus-Merzbacher Disease. Stem Cells. 2016;34(4):984-996.
[36] MAGNANI D, CHANDRAN S, WYLLIE DJA, et al. In Vitro Generation and Electrophysiological Characterization of OPCs and Oligodendrocytes from Human Pluripotent Stem Cells. Methods Mol Biol. 2019;1936: 65-77.
[37] PRASAD A, TEH DBL, BLASIAK A, et al. Static Magnetic Field Stimulation Enhances Oligodendrocyte Differentiation and Secretion of Neurotrophic Factors. Sci Rep. 2017;7(1): 6743.
[38] WANG C, ZHANG CJ, MARTIN BN, et al. IL-17 induced NOTCH1 activation in oligodendrocyte progenitor cells enhances proliferation and inflammatory gene expression. Nat Commun. 2017;8:15508.
[39] LIM CT, ZHOU EH, QUEK ST. Mechanical models for living cells--a review. J Biomech. 2006;39(2):195-216.
[40] JEN CJ, JHIANG SJ, CHEN HI. Invited review: effects of flow on vascular endothelial intracellular calcium signaling of rat aortas ex vivo. J Appl Physiol (1985). 2000;89(4):1657-1662, 1656.
[41] BACHRACH NM, VALHMU WB, STAZZONE E, et al. Changes in proteoglycan synthesis of chondrocytes in articular cartilage are associated with the time-dependent changes in their mechanical environment. J Biomech. 1995;28(12):1561-1569.
[42] LIU SQ. Influence of tensile strain on smooth muscle cell orientation in rat blood vessels. J Biomech Eng. 1998;120(3): 313-320.
[43] ZHANG C, ZHOU L, ZHANG F, et al. Mechanical remodeling of normally sized mammalian cells under a gravity vector. FASEB J. 2017;31(2): 802-813.
[44] 吕东媛,周吕文,龙勉.干细胞的生物力学研究[J].力学进展, 2017, 47(1): 534-585.
[45] WANG CY, DENEEN B, TZENG SF. BRCA1/BRCA2- containing complex subunit 3 controls oligodendrocyte differentiation by dynamically regulating lysine 63-linked ubiquitination. Glia. 2019;67(9):1775-1792.
[46] KAINDL J, MEISER I, MAJER J, et al. Zooming in on Cryopreservation of hiPSCs and Neural Derivatives: A Dual-Center Study Using Adherent Vitrification. Stem Cells Transl Med. 2019;8(3):247-259.