Hybridoma Cryoprotective Media (HCM); DMSO Free

Description

Background

PurMa^TM Hybridoma Cryoprotective Media (HCM); DMSO Free is an excellent choice for cryopreservation of hybridoma cell lines. We strongly recommend applying DMSO free cryoprotective media only for cell lines with sensitivity to DMSO.  PurMa^TM HCM has great performance in preservation of fused myeloma and CHO cells (producing antibodies) as well as humanized Hybridomas. It contains a proprietary combination for the best hybridoma cryopreservation, survival, and recovery after thawing.

DMSO benefits cryoprotection and cryoprotective agent in several ways

1. Provides strong molecular stability.
2. Besides, facilitates a high non-conductor constant.
3. It contains the basic properties, salts solubility particularly for anions. Additionally contains a propensity to act as the acceptor atom in hydrogen bonding.
4. Especially,  DMSO makes a  hydrogen bond with water which is 30% stronger than between two water molecules.
5. The proprietary formulation of PurMa^TM Mammalian Cryoprotective Media (MCM) allows the MCM to get inside the cells instantly and lower cell death because of penetration/freezing.

Although, DMSO with the penetrating ability, is a strong cryoprotective agent, there is however some limitations and disadvantages. For more details about the beneficial role of DMSO,  please see our other cryoprotective media with DMSO, such as:

limitations and disadvantages of DMSO

1. Basically, it has a broad toxicity.
2. Additionally, it modifies the expression of cells such an NK and T cells
3. It alters the mRNA level of the de novo DNA methyltransferase DNMT3A. This, therefore, is accompanied by hyper- or hypo-methylation of many genetic loci.
4. Moreover, it causes difficulties in scaling up DMSO-based cryoprotectants.
5. It damages the mitochondria in certain cell culture models such as astrocytes.
6. It negatively impacts cellular membrane/cytoskeleton structure integrity by interacting with proteins and dehydrating lipids.
7. Besides, it induces unwanted stem cell differentiation.
8. Affects cellular epigenetic profile resulting in undesirable phenotypic disturbances. That is basically because of repeated usage of DMSO even at sub-toxic levels.
9. Afterwards, it is necessary to remove DMSO by intensive washing. That in turn results in cell sensitivity to osmotic/mechanical stresses occurring. This is a costly and time-consuming process during freezing and thawing, and moreover leads to a significant loss of cell number.

PurMa^TM Hybridoma Cryoprotective Media (HCM); DMSO Free provides high post-thaw cell viability. Besides, it preserves morphology, proliferation as well as  the differentiation potentials of hybridoma cells.

Special Features

• Ready-to-Use medium.
• It does not contain DMSO or any other penetrating agents.
• Each Lot of PurMa^TM HCM goes through pathogens and sterility evaluations.
• Additionally, we perform the following survival tests on each Lot of PurMa^TM HCM:  Survival test (by Trypan Blue method), viability & performance evaluation (by MTT Test), and finally,  for the rate of recovery by Alamar Blue test.
• Moreover, PurMa^TM HCM has been designed for the lowest cell loss during freezing and thawing.
• PurMa^TM HCM yields highest cell recovery, 64% survival and recovery rate after long term cryopreservation.
• PurMa^TM HCM gives faster recovery which results in less exposure to the toxins from the non-recovering cells.
• Preserves morphological appearance as the main recovery factor for cells.
• Tested for pH, osmolality, sterility, and endotoxin.
• PurMa^TM HCM has been tested successfully for several antibody making cell lines including fused NS1 and CHO cells.
• Suitable for clinical applications.

Guideline

for PurMa Biologics guideline for cryoprotection, click here as well as in corresponding tab: PurMa™ Guideline for Cryoprotecting various cell lines

References

1. Differentiation of erythroleukemic cells in vitro: irreversible induction by dimethyl sulfoxide (DMSO). Preisler et al. 1975 Jun;85(3):537-46. doi: 10.1002/jcp.1040850305.
2. Dimethyl sulfoxide-free cryopreservation solutions for hematopoietic stem cell grafts. Kaushal et al. 2022 Mar;24(3):272-281. doi: 10.1016/j.jcyt.2021.09.002. Epub 2021 Oct 12.

Learn more: What Is Cryoprotective Media?