Comparison of Microglial Morphology and Function in Primary Cerebellar Cell Cultures on Collagen and Collagen-Mimetic Hydrogels

被引:5
作者
Balion, Zbigniev [1 ,2 ]
Svirskiene, Natasa [3 ]
Svirskis, Gytis [3 ]
Inokaitis, Hermanas [4 ]
Cepla, Vytautas [5 ,6 ]
Ulcinas, Arturas [6 ]
Jelinskas, Tadas [5 ,6 ]
Eimont, Romuald [5 ,6 ]
Pauziene, Neringa [4 ]
Valiokas, Ramunas [5 ,6 ]
Jekabsone, Aiste [1 ,7 ]
机构
[1] Lithuanian Univ Hlth Sci, Fac Pharm, Inst Pharmaceut Technol, Lab Pharmaceut Sci, Sukilelitu Ave 13, LT-50162 Kaunas, Lithuania
[2] Lithuanian Univ Hlth Sci, Neurosci Inst, Lab Biochem, Eiveniu Str 4, LT-50161 Kaunas, Lithuania
[3] Lithuanian Univ Hlth Sci, Neurosci Inst, Lab Neurophysiol, Eiveniu Str 4, LT-50161 Kaunas, Lithuania
[4] Lithuanian Univ Hlth Sci, Fac Med, Inst Anat, Mickeviciaus 9, LT-43074 Kaunas, Lithuania
[5] Ferentis UAB, Savanoriu 231, LT-02300 Vilnius, Lithuania
[6] Ctr Phys Sci & Technol, Dept Nanoengn, Savanoriu 231, LT-02300 Vilnius, Lithuania
[7] Lithuanian Univ Hlth Sci, Inst Cardiol, Lab Preclin Drug Invest, Sukileliu Ave 15, LT-50162 Kaunas, Lithuania
关键词
collagen-like peptide; crosslinked collagen; hydrogel; TEM; cryo-FIB/SEM; microglia; Ca2+ oscillations; ORGANOIDS; BRAIN; GENERATION; TISSUE;
D O I
10.3390/biomedicines10051023
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Neuronal-glial cell cultures are usually grown attached to or encapsulated in an adhesive environment as evenly distributed networks lacking tissue-like cell density, organization and morphology. In such cultures, microglia have activated amoeboid morphology and do not display extended and intensively branched processes characteristic of the ramified tissue microglia. We have recently described self-assembling functional cerebellar organoids promoted by hydrogels containing collagen-like peptides (CLPs) conjugated to a polyethylene glycol (PEG) core. Spontaneous neuronal activity was accompanied by changes in the microglial morphology and behavior, suggesting the cells might play an essential role in forming the functional neuronal networks in response to the peptide signalling. The present study examines microglial cell morphology and function in cerebellar cell organoid cultures on CLP-PEG hydrogels and compares them to the cultures on crosslinked collagen hydrogels of similar elastomechanical properties. Material characterization suggested more expressed fibril orientation and denser packaging in crosslinked collagen than CLP-PEG. However, CLP-PEG promoted a significantly higher microglial motility (determined by time-lapse imaging) accompanied by highly diverse morphology including the ramified (brightfield and confocal microscopy), more active Ca2+ signalling (intracellular Ca2+ fluorescence recordings), and moderate inflammatory cytokine level (ELISA). On the contrary, on the collagen hydrogels, microglial cells were significantly less active and mostly round-shaped. In addition, the latter hydrogels did not support the neuron synaptic activity. Our findings indicate that the synthetic CLP-PEG hydrogels ensure more tissue-like microglial morphology, motility, and function than the crosslinked collagen substrates.
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页数:19
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