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Cell freezing medium

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Cell freezing medium is a specialized liquid solution designed to protect cells during the freezing process. It helps to maintain the viability and integrity of cells by preventing damage from ice crystal formation and osmotic stress. This product is intended for use in cryopreservation of various cell types.

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Lab products found in correlation

Penicillin, by Thermo Fisher Scientific (3 mentions) Streptomycin, by Thermo Fisher Scientific (3 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Dulbecco s modified eagle medium, by Thermo Fisher Scientific (2 mentions) Act diff, by Beckman Coulter (1 mentions) Amphotericin b, by Thermo Fisher Scientific (1 mentions) Trypsin edta, by Merck Group (1 mentions) Fungizone, by Thermo Fisher Scientific (1 mentions) Alpha mem, by STEMCELL (1 mentions)

Spelling variants of this product

Recovery Cell Culture Freezing Medium (88 citations) Freezing medium (7 citations) Cell Culture Freezing Medium (3 citations) Gibco Cell Culture Freezing Medium (2 citations) Recovery cell freezing medium (2 citations)

4 protocols using cell freezing medium

1

Isolation and Cryopreservation of PBMCs

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Separated plasma was aliquoted and stored at −20 °C. The extracted PBMCs were frozen in cell freezing medium (Gibco) and stored in a deep freezer (−80 °C). The number of total cells was counted, and the relative proportions of cell populations was determined by flow cytometry using surface markers for T cells, B cells, NK cells, and monocytes.
Suojalehto H., Ndika J., Lindström I., Airaksinen L., Karvala K., Kauppi P., Lauerma A., Toppila-Salmi S., Karisola P, & Alenius H. (2021). Transcriptomic Profiling of Adult-Onset Asthma Related to Damp and Moldy Buildings and Idiopathic Environmental Intolerance. International Journal of Molecular Sciences, 22(19), 10679.
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2

PBMC Isolation and Characterization

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Peripheral blood mononuclear cells (PBMCs) were isolated from 8 ml of whole blood by a CPT tube system (BD Sciences). The separated plasma was aliquoted and stored at -20°C. The extracted PBMCs were frozen in cell-freezing medium (Gibco) and stored in a deep freezer (-80°C). After thawing the samples, the number of total cells was counted (Beckman Coulter AcT/Diff), and the relative proportions of cell populations were determined by flow cytometry using surface markers. Subsets of T cells (CD3+), B cells (CD19+), NK cells (CD16+ CD56+), and monocytes (CD14+) were identified. In addition, cell viability was checked, and the percentage of dead cells was determined.
Karisola P., Palosuo K., Hinkkanen V., Wisgrill L., Savinko T., Fyhrquist N., Alenius H, & Mäkelä M.J. (2021). Integrative Transcriptomics Reveals Activation of Innate Immune Responses and Inhibition of Inflammation During Oral Immunotherapy for Egg Allergy in Children. Frontiers in Immunology, 12, 704633.
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3

Isolation and Culture of hMSCs

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hMSCs were isolated from fresh bone marrow aspirate purchased from Lonza (donor 18-year-old black female). Following previously published protocols13 (link),22 , hMSCs were isolated based on preferential adhesion to tissue culture polystyrene plates. Freshly isolated hMSCs were detached with 0.05% trypsin–EDTA (Sigma) and subsequently centrifuged, counted, and frozen in Cell Freezing Medium (Thermo Fisher). Only passage 2 or 3 cells were used for all encapsulation experiments. Growth media consisted of low glucose (1 ng/mL glucose) Dulbecco’s Modified Eagle Medium (ThermoFisher) supplemented with 10% FBS (ThermoFisher), 1 ng/ml fibroblast growth factor basic (bFGF) (Life Technologies), 50 U/ml penicillin (ThermoFisher), 50 μg/ml streptomycin (ThermoFisher), 0.5μg/ml of Amphotericin B (ThermoFisher). For secretion experiments, the same media was used sans bFGF (referred to as Experimental Media).
Caldwell A.S., Rao V.V., Golden A.C, & Anseth K.S. (2019). Porous bio-click microgel scaffolds control hMSC interactions and promote their secretory properties. Biomaterials, 232, 119725.
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4

Isolation and Expansion of Rat Mesenchymal Stem Cells

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Immediately after euthanasia, rMSCs were isolated from the bone marrow of the humerus of each rat. Briefly, the humerii from each rat was removed, the ends of the bone cut, and the bone marrow flushed with growth media and plated directly onto tissue culture plates. rMSCs were cultured in alpha modified Eagle medium (alpha-MEM, Stem Cell Technologies) with nucleosides supplemented with 50 μg/mL penicillin (Gibco), 50 μg/mL streptomycin (Gibco), and 1 μg/mL fungizone (Gibco) containing 10% (vol/vol) fetal bovine serum (Gibco). Media was changed every 2-3 days. After 10 days (70-80% confluency), rMSCs from each individual rat were frozen down in a solution of 40% growth media, 40% FBS and 20% DMSO. Passage 1 (P1) rMSCs from individual rats were pooled, expanded for an additional 7-10 days, and frozen at P2 in Cell Freezing Medium (ThermoFisher). All experiments used P3 pooled SHAM and OVX rMSCs.
Rao V.V., Wechsler M.E., Cravens E., Wojda S.J., Caldwell A.S., Kirkpatrick B.E., Donahue S.W, & Anseth K.S. (2022). Granular PEG hydrogels mediate osteoporotic MSC clustering via N-cadherin influencing the pro-resorptive bias of their secretory profile. Acta biomaterialia, 145, 77-87.
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