Process for preserving insulin-secreting cells intended to be transplanted in a patient
Process for preserving insulin secreting cells intended to be transplanted, including the following steps: introducing an initial volume of culture medium and insulin-secreting cells into a culture container, providing a culture medium source, and replacing, at time intervals below 8 hours, the culture medium in the culture container with culture medium from the source so as to renew the culture medium contained in the container.
1. A process for preserving pancreatic islets intended to be transplanted, including the following steps:
introducing an initial volume of culture medium and pancreatic islets into a culture container, said pancreatic islets being contained in a preparation comprising exocrine tissue that releases enzymes with concentrations that are detrimental to the pancreatic islets,
providing a culture medium source having new culture medium,
replacing, at time intervals below 8 hours, the culture medium in said culture container with said new culture medium from said source so as to renew the culture medium contained in said container, so that the enzymes released by exocrine tissue are removed from the culture container,
forming the culture container by a flexible pouch preventing adhesion of the pancreatic islets,
providing the culture container with at least one deflector arranged on a direct flow path between an inlet orifice and an outlet orifice of the culture container, and
providing said deflector in the form of a weld extending perpendicularly from a side on which said inlet and outlet orifices are arranged, said weld being formed between said inlet and outlet orifices.
2. The process according to claim 1 , further comprising introducing a concentration of the pancreatic islets in the culture medium between 400 and 40,000 islet equivalents/mL.
3. The process according to claim 1 , wherein said culture medium introducing step comprises introducing a culture medium which includes a base medium supplemented with albumin.
4. The process according to claim 1 , wherein said replacing step comprises the culture medium contained in the container every 2 hours so as to periodically renew the culture medium contained in said container.
5. The process according to claim 1 , further comprising performing the culture medium replacing step by establishing a continuous flow of culture medium coming from the medium source, in which said flow is supplied to the culture container by means of the at least one inlet orifice and is removed from said culture container by means of the at least one outlet orifice, so as to continuously renew the culture medium contained in said container.
6. The process according to claim 5 , further comprising maintaining a stationary phase in which the flow is arranged to keep the volume of culture medium contained in the culture container constant.
7. The process according to claim 6 , further comprising prior to the stationary phase, arranging the flow to increase the initial volume of culture medium.
8. The process according to claim 5 , further comprising the step of establishing the continuous flow of culture medium comprises establishing the flow in a period greater than 24 hours.
9. The process according to claim 5 , further comprising the step of establishing the continuous flow of culture medium comprises establishing the flow in a period between 24 hours and 72 hours.
10. The process according to claim 5 , further comprising providing a flow rate of the flow supplying the at least one inlet orifice between 0.2 and 5 mL/minute.
11. The process according to claim 5 , further comprising oxygenating the flow of culture medium before being supplied to the at least one inlet orifice.
12. The process according to claim 5 , further comprising providing at least some of the flow of culture medium supplying the at least one inlet orifice from the flow of culture medium that has previously been removed through the at least one outlet orifice.
13. The process according to claim 5 , further comprising connecting the at least one inlet orifice of the culture container by means of a first tubing to a container forming a culture medium source, in which a first peristaltic pump head is associated with said first tubing so as to supply the at least one inlet orifice with the flow of culture medium coming from said source container.
14. The process according to claim 13 , further comprising connecting the at least one outlet orifice of the culture container by means of a second tubing to a container intended to contain the culture medium removed through the at least one outlet orifice, a second peristaltic pump head being associated with the second tubing so as to remove, from the container, the flow of culture medium through the at least one outlet orifice.
15. The process according to claim 14 , further comprising connecting a third tubing between the first and second tubings, so that at least some of the flow flowing into the second tubing is reintroduced into the first tubing.
16. The process according to claim 13 , further comprising placing a system for aeration of the flow of culture medium on the flow path of the first tubing.