Delivery of high cell mass in a syringe and related methods of cryopreserving cells

A method of rapidly freezing cells, comprising:

• acquiring a desired quantity of cells for cryostorage;
• suspending the acquired cells in chilled freeze media containing a permeating cryoprotectant, wherein the freeze media is at a temperature of approximately 0° C. to 4° C.;
• placing the cells and freeze media in an apparatus configured to store and dispense cryopreserved cells, wherein at least a portion of the apparatus is made from a biocompatible material;
• and
• rapidly cooling the apparatus containing the cells and chilled freeze media to a temperature of −130° C. or below at a rate of approximately 8° C./minute.

A method for cryostoring cells comprising:

• acquiring a desired quantity of cells for cryostorage;
• placing the acquired cells in chilled freeze media containing a permeating cryoprotectant;
• storing the cells and freeze media in an apparatus suitable for cryostorage, wherein the apparatus is configured to store and dispense cryopreserved cells and comprises:
• a body having an open first end and an open second end;
• a first cap configured to removably attach to the open first end;
• a second cap configured to removably attach to the open second end;
• a plunger portion contained within the body and adjacent to one of said open ends;
• and
• a plunger rod configured to be connected to the plunger portion, wherein at least a portion of the apparatus is made from a biocompatible material;
• and
• cooling the apparatus to an approximate temperature of −130° C. or below.

A method for inoculating a bioreactor with cryopreserved cells comprising:

• acquiring a desired quantity of cells for cryostorage;
• placing the acquired cells in chilled freeze media containing a permeating cryoprotectant;
• storing the cells and freeze media in an apparatus configured to store and dispense cryopreserved cells, wherein the apparatus comprises:
• a body having an open first end and an open second end;
• a first cap configured to removably attach to the open first end;
• a second cap configured to removably attach to the open second end;
• a plunger portion contained within the body and adjacent to one of said open ends;
• and
• a plunger rod configured to be connected to the plunger portion, wherein at least a portion of the apparatus is made from a biocompatible material;
• cooling the apparatus to an approximate temperature of −130° C. or below;
• subsequent to cooling the apparatus to an approximate temperature of −130° C. or below, transferring the frozen media and cells from the apparatus to a thawing vessel containing growth media at a temperature substantially warmer than 0° C.;
• and
• inoculating a bioreactor with the cells from the thawing vessel.

A method of freezing a large cell mass at a high density, comprising:

• suspending a large cell mass in a freeze media containing a permeating cryoprotectant, wherein the concentration of the permeating cryoprotectant is sufficient to permit the cells to be stored at a density greater than 1.5×108 cells/ml;
• placing the cells and freeze media in a storage apparatus;
• and
• cooling the cells and freeze media to a temperature at or below approximately −130° C.

A method of freezing a large cell mass at a high density, comprising:

• suspending a large cell mass in a freeze media containing 20% DMSO, wherein the freeze media does not include animal serum and wherein the concentration of the DMSO is sufficient to permit the cells to be stored at a density greater than 3.0×107 cells/ml;
• placing the cells and freeze media in a storage apparatus;
• and
• cooling the cells and freeze media to a temperature at or below approximately −130° C.