Home

Description

Search Catalogues

Browse catalogues

Collections

Guidelines

Search Web Site

Contacts

FAQ

Site Map

Mirrors

LABORATORY PROCEDURES FOR PLANT CELL LINES

REFERENCE NO.: PC/1998/3/4


TITLE: ROUTINE MAINTENANCE AND STORAGE OF CELL LINES - FROZEN CELL LINES -


INTRODUCTION

If a cell line passed the preliminary quality checks (PC/1998/2/2PC/1998/2/3) it is further propagated to check whether the given cryopreservation procedure can be carried out at the collection centre. Furthermore it is tested whether the regrowing samples can be considered to be identical with the original cell line. If no differences between the initial cell line and the regrowing frozen ones can be detected or if the depositor decides that the regrowing samples are identical or identical with respect to a desired trait 40 ampoules are cryopreserved (PC/1998/3/2), 20 of which are stored in different containers. One of the containers represents the "Master Collection", the other one the "Working Collection".

In special cases, if a cell line is requested very frequently, it can be decided that more samples are stored in the "Master-" and/or the "Working Collection".

PROCEDURE

1. Before storage colour code buttons have to be inserted into the lid of the cryovials so that the colour differs from those used for cryovials of other cell lines placed next to the new one in the desired position in the cryocontainer.

2. 20 ampoules are stored in the "Master Collection" (Container 1)

3. 20 ampoules are stored in the "Working Collection" (Container 2)

4. The position of the samples is entered into the computer database (PC/1998/3/4.1) for cryocontainer management.

5. A printout is made for those trays in the crypcontainer in which new samples have been placed. The date is stamped on this printout and it is stored in the cryocontainer document file (book) (PC/1998/3/4.2).

6. On request of a plant cell line the percentage of regrowth for the last series of samples has to be looked up.

7.1. If the percentage of recovery is higher than 50 %, 3 samples from the "Working Collection" are recovered and sent to the client (PC/1998/4/1).

7.2. If the percentage of recovery is lower than 50 %, 5 samples from the "Working Collection" are recovered and sent to the client (PC/1998/4/1)

8. If samples are taken out of any of the cryocontainers this is recorded in the computer data base (PC/1998/3/4.1) for cryocontainer management and a printout is made of the for the tray concerned. The date is stamped on the printout and it is stored in the cryocontainer document file (book) (PC/1998/3/4.2).

9. If the last sample from the "Working Collection" has been used an adequate number of samples from the "Master Collection" is recovered (3 samples for those cultures with a percentage of recovery higher than 50 % and 5 samples for cell lines with a percentage of recovery lower than 50 %) and a new series of samples is cryopreserved for the "Working Collection".

10. Five ampoules are thawed, regrown and checked for identity with the original cell lines. If no differences between the initial cell line and the regrown frozen ones can be detected or if the depositor decides that the regrowing samples are identical or identical with respect to a desired trait, the cryopreserved ampoules are integrated into the "Working Collection".

11. The position of the samples is entered into the computer data base (PC/1998/3/4.1) for cryocontainer management.

12. A printout is made for those trays in the cryocontainer into which new samples have been placed. The date is stamped on this printout and it is stored in the cryocontainer document file (book) (PC/1998/3/4.2).

13. If only 5 frozen ampoules are left in the "Master Collection", an adaequate number of samples from the "Master Collection" is recovered (3 samples for those cultures with a percentage of recovery higher than 50 % and 5 samples for cell lines with a percentage of recovery lower than 50 %) and a new series of samples is cryopreserved for the "Master Collection".

14. Five ampoules are thawed, regrown and checked for identity compared to the original cell lines. If no differences between the initial cell line and the regrowing frozen ones can be detected or if the depositor decides that the regrown samples are identical or identical with respect to a desired trait the cryopreserved ampoules are integrated into the "Master Collection".

15. The position of the samples is entered into the computer data base (PC/1998/3/4.1) for cryocontainer management.

16. A printout is made for those trays in the cryocontainer into which new samples have been placed. The date is stamped on this printout and it is stored in the cryocontainer document file (book) (PC/1998/3/4.2).


Guidelines prepared for CABRI by DSMZ, 20 Jan. 1998
Page Layout by CERDIC
Copyright CABRI, 1998 

© The CABRI Consortium 1999-2013.
This work cannot be reproduced in whole or in part without the express written permission of the CABRI consortium.
Site maintained by Paolo Romano. Last revised on April 2013.