TY - JOUR

T1 - Cryopreservation of cells using defined serum-free cryoprotective agents

AU - Volbers, Jan Cedric

AU - Lauterböck, Lothar

AU - Hofmann, Nicola

AU - Glasmacher, Birgit

N1 - Funding Information: Acknowledgment: The authors want to thank Julia Struß for her technical support. Also the authors acknowledge Thomas Müller for kindly donating the multipotent stromal cells from the common marmoset monkey (Callithtix jacchus). This work has been partially supported by funding from the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) for the Cluster of Excellence REBIRTH (From Regenerative Biology to Reconstructive Therapy) (EXC 62/1) and ZIM (Zentrales Innovationspro-gramm Mittelstand, KF2654703SB3) in cooperation with Askion.

PY - 2016/9

Y1 - 2016/9

N2 - For regenerative purposes, there is a high demand for viable and active cells. A big issue is to have enough viable cells available at any given time. One solution is cryopreservation. In this context, DMSO is used as cryoprotective agent (CPA) along with fetal bovine serum for nutrient supply and stress shielding effects. To use these cells for human clinical studies, it is important to eliminate the serum to prevent foreign immune reactions and virus transmittance and DMSO for its toxic effect. In this study a serum free cryopreservation solution and protocol has been established. The combination of methylcellulose and poloxamer 188 provide the basis for the new CPA. Other additves are -tocopherol, ectoine, prolin and ascorbic acid. The CPAs were examined with 3T3-cells and multipotent stromal cells from the common marmoset monkey (Callithrix jacchus). The cells were preserved with various CPA concentrations, incubation times and different cooling rates. To enable a higher throughput of encouraging conditions a fluorescence microscopy analysis was used. The use of methylcellulose, poloxamer 188 and -tocopherol enables the reduction of DMSO [up to 2.5% (v/v)] and the elimination of serum without viability losses compared to control.

AB - For regenerative purposes, there is a high demand for viable and active cells. A big issue is to have enough viable cells available at any given time. One solution is cryopreservation. In this context, DMSO is used as cryoprotective agent (CPA) along with fetal bovine serum for nutrient supply and stress shielding effects. To use these cells for human clinical studies, it is important to eliminate the serum to prevent foreign immune reactions and virus transmittance and DMSO for its toxic effect. In this study a serum free cryopreservation solution and protocol has been established. The combination of methylcellulose and poloxamer 188 provide the basis for the new CPA. Other additves are -tocopherol, ectoine, prolin and ascorbic acid. The CPAs were examined with 3T3-cells and multipotent stromal cells from the common marmoset monkey (Callithrix jacchus). The cells were preserved with various CPA concentrations, incubation times and different cooling rates. To enable a higher throughput of encouraging conditions a fluorescence microscopy analysis was used. The use of methylcellulose, poloxamer 188 and -tocopherol enables the reduction of DMSO [up to 2.5% (v/v)] and the elimination of serum without viability losses compared to control.

KW - Antioxidants

KW - Cryopreservation

KW - Fluorescence microscopy

KW - Methylcellulose

KW - Multipotent stromal cells

KW - Poloxamer 188

KW - Serum free

UR - http://www.scopus.com/inward/record.url?scp=85059823520&partnerID=8YFLogxK

U2 - 10.1515/cdbme-2016-0070

DO - 10.1515/cdbme-2016-0070

M3 - Article

AN - SCOPUS:85059823520

VL - 2

SP - 315

EP - 318

JO - Current Directions in Biomedical Engineering

JF - Current Directions in Biomedical Engineering

IS - 1

ER -