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LABORATORY PROCEDURES FOR PLANT CELL LINES

REFERENCE NO.: PC/1998/3/1


 TITLE: QUALITY CONTROL, MAINTENANCE, PRESERVATION, STORAGE AND ACCEPTANCE OF CELL LINES - LIVING CULTURES -


INTRODUCTION

If a cell line has passed the preliminary quality tests it is further propagated to check whether cultivation is successfully carried out over three transfer cycles and to produce material for the final quality checks and authentication. When the final quality check and authentication have been carried out and the cultivation conditions are acceptable for the collection centre the cell line is finally accepted and included into the public collection.

PROCEDURE

1. Cells are propagated until enough material has been produced for characterisation or for at least three transfer cycles.

1.1 Cells are propagated as described by the depositor until enough material has been produced for characterisation or for at least three transfer cycles.

1.2. If the cultivation conditions given by the depositor differ from standard conditions used in CABRI and are not acceptable for maintenance of cultures in the long term the cell line is in addition cultivated by standard procedures for callus or suspension (PC/1998/3/3) which mimic the given conditions most.

1.3. Material is sent back to the depositor. The depositor is asked to decide whether the cell line may be further propagated under CABRI standard conditions on the collection standard media (PC/1998/3/1 Appendix 1).

1.4 If the depositor accepts further maintenance of the cell line under CABRI standard conditions, these conditions are used to produce enough cell material for further quality control and authentication. If not, the accession procedure is halted and the depositor informed (PC/1998/3/1 Appendix 2)

2. Growth and morphological characters are collected by standard procedures (PC/1998/3/1.1) and stored in the database.

3. 2 g of callus or suspension cells (fresh weight) are extracted with methanol according to the standard procedure (PC/1998/3/1.2)

3.1 The extracts are analysed with HPLC (PC/1998/3/1.3) and the HPLC profiles are stored in a database.

3.2. Additionally UV spectra of the unpurified extracts may be measured and stored in a data base (PC/1998/3/1.4)

4. After quality control and authentication the cell line is finally accepted and further maintained by the standard procedures for cell culture management (PC/1998/3/3). All data are entered into the plant cell culture database.


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

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