Klinische Wochenschrift

, Volume 65, Issue 11, pp 507–512 | Cite as

Establishment of primary cell cultures: Experiences with 155 cell strains

  • M. Dietel
  • H. Arps
  • D. Gerding
  • M. Trapp
  • A. Niendorf
Originalien

Summary

Cell culture systems allow the examination of cell populations in a functional state. To simulate in vivo conditions as closely as possible freshly established cell strains are superior to permanent cell lines. Different aspects for the establishment of primary cell cultures obtained from various tissues are compared: (1) Disintegration, (2) culture media supplemented with basal additions, (3) special supplements (growth factors, hormones), and (4) attachment factors. The proliferation rates of the attained cell strains were evaluated by determination of cell doubling times. Procedures for how to obtain a relatively high plating efficiency (approx. 70% in our series of 219 attempts) of primary growth in vitro are described: (1) Mechanical disintegration is superior to enzymatic digestion. If mechanical treatment alone did not produce a sufficient number of viable cells, additional digestion with collagenase/dispase revealed a higher number of proliferating primary cultures than with trypsin. (2) Proliferation of cell cultures from normal and tumorous tissues of epithelial origin was superior in Leibovitz L 15 medium (58 of 87 (67%) cases). Cultures from mesenchymal tissues and tumors were found to have shortest cell doubling times in MEM and RPMI 1640 (16 of 23 (70%) cases). The media were supplemented with the basal additions indicated. (3) In approx. 30% of the cases special supplements like growth factors or hormones increased cell replication, although they were almost always not essential for cell growth. (4) Attachment factors only rarely contributed to the initiation of primary monolayer cultures. The application of various culture conditions does not lead to a protocol optimal for all tissues, for all probes of the same type of tumor, or for all tumor specimens of unique differentiation.

Key words

Cell culture techniques Primary cell cultures Growth factors Culture media Attachment factors 

Abbreviations

EGF

epidermal growth factor

FGF

fibroblast growth factor

GHL

glycyl-L-histidyl-L-lysine

PAN

pancreozymin

E

estrogen

PR

progesterone

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Copyright information

© Springer-Verlag 1987

Authors and Affiliations

  • M. Dietel
    • 1
  • H. Arps
    • 1
  • D. Gerding
    • 1
  • M. Trapp
    • 1
  • A. Niendorf
    • 1
  1. 1.Institut für Pathologie, U.K. EppendorfUniversität HamburgGermany

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