Giovarelli 2023 Mol Med

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Giovarelli M, Serati A, Zecchini S, Guelfi F, Clementi E, MandΓ² C (2023) Cryopreserved placental biopsies maintain mitochondrial activity for high-resolution respirometry.

Β» Mol Med 29:45. PMID: 37013473 Open Access

Giovarelli Matteo, Serati Anais, Zecchini Silvia, Guelfi Fabiola, Clementi Emilio, Mando Chiara (2023) Mol Med

Abstract: High-resolution respirometry (HRR) of human biopsies can provide useful metabolic, diagnostic, and mechanistic insights for clinical research and comparative medical studies. Fresh tissues analysis offers the potential best condition, the drawback being the need to use them shortly after dissection for mitochondrial respiratory experiments. The development of effective long-term storage protocols for biopsies that allow the assessment of key Electron Transport System (ETS) parameters at later stages is thus a major need.

We optimised a cryopreservation protocol that preserves mitochondrial membranes intactness, otherwise affected by direct tissue freezing. The protocol is based on a gradual freezing step from on-ice to liquid nitrogen and - 80 Β°C storage using a specific DMSO-based buffer.

Placenta is a suitable tissue to design and test the effectiveness of long-term storage protocols being metabolically active foetal tissue with mitochondrial dysfunctions contributing to placental disease and gestational disorders. Here we designed and tested the effectiveness of the cryopreservation protocol using human placenta biopsies; we measured the ETS activity by HRR of placenta specimens comparing fresh, cryopreserved, and snap frozen conditions.

By this protocol, Oxygen Consumption Rate (OCR) measurements of fresh and cryopreserved placental specimens are comparable whereas snap frozen procedure impairs mitochondrial activity. β€’ Keywords: Cryopreservation, High-resolution respirometry, Mitochondria, Placental biopsies β€’ Bioblast editor: Plangger M β€’ O2k-Network Lab: IT Milan Clementi E

Labels: MiParea: Respiration, Instruments;methods 

Stress:Cryopreservation  Organism: Human  Tissue;cell: Genital  Preparation: Permeabilized tissue 

Coupling state: LEAK, OXPHOS, ET  Pathway: N, S, CIV, NS, ROX  HRR: Oxygraph-2k 


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