Instructions for Cryopreservation

Introduction

Cryopreservation may compromise cell quality and performance. Performance of the cells CANNOT be guaranteed after cryopreservation. These instructions do not apply to Clonetics Hepatocytes, Neural Progenitor Cells, Poietics Adipose-Derived Stem Cells or Bovine Brain Microvascular Endothelial Cells.

Cryo freezing solutions used for Clonetics and Poietics products:

Clonetics Cells (see exceptions below)

80% Clonetics Growth Medium of Choice

10% FBS

10% DMSO

Mesenchymal Stem Cells

70% MSCBM

10% DMSO

20% Human Serum Albumin

Skeletal Muscle Cells

70% SkGM

20% FBS

10% DMSO

Poietics Cells

86.5% IMDM

7.5% DMSO

4% Human Serum Albumin

2% Hydroxy-ethyl-starch

General Instructions

1. Sterile filter freezing media using any 0.2 micron filter.

2. Harvest cells and centrifuge to collect cells into a pellet.

3. Resuspend cells in cold freezing solution at 500,000 to 2,000,000 cells per ml. Work quickly! Once exposed to DMSO, cells become very fragile.

4. Pipet aliquots (1 ml each) into freezing vials or ampoules and seal.

5. Insulate aliquots with Styrofoam or propanol freezing canister.

6. Store cells at -70°C overnight.

7. Within 12 to 24 hours, place in liquid Nitrogen for long-term storage (-200°C). Cells will be compromised by storage in -70°C.

Note: Lonza CANNOT guarantee performance of Clonetics and Poietics Cells that have been cryopreserved. To avoid loss of cells and forfeiture of your warranty, we recommend keeping cells in continuous culture without cryopreservation.