Here I summarize key takeaways and implications from a paper that describes the establishment of the only aquatic invertebrate cell line. The author is Eder L. Hansen.
Hansen (1976) Hansen EL. A cell line from embryos of Biomphalaria glabrata Pulmonata Establishment and characteristics. Academic Press; Cambridge: 1976. Google Scholar.
Paper Summary
This article describes the establishment and characteristics of the one of the only aquatic invertebrate cell line available to date. Hansen was able to derive a cell line from embryonic cells of a freshwater snail through spontaneous immortalization. Specifically, the author began cultured Bge (Biomphalaria glabrata embryo) 740 primary cultures over the course of a 2-year period through an array of methods. They describe Bge primary cultures as being tolerant to a wide variety of culturing media which made it initially difficult in determining an optimum method. Furthermore, they utilized existing information to supplement the media with the appropriate organic osmolytes like galactose. However, they determined that any range of compositions of the media could work as long as it was in an appropriate osmolality and pH ranges. The medium used for the primary culture that eventually went on to become a cell line was one of a simpler composition. Furthermore, they claim that early division of primary cultures, a technique that differs from those usually employed, should be utilized for other invertebrate tissue sources. By this they simply mean the early trypsinization of the primary cultures to disassociate and passage cells was the key to the selection method which led to their cell line.
So what?
Although the organism they use is a freshwater invertebrate and not a marine invertebrate like my model organism Botryllus schlosseri there is still key approaches presented in Hansen 1976 which should be taken into consideration when approached the development of a cell line for my model organism.
Particularly, what caught my attention was:
The early integration of trypsinization of primary cultures.
The tolerance of a wide range of culturing conditions for Bge primary cultures as early work on Botryllus schlosseri somatic primary cultures also demonstrated a similar tolerance.
The brief immersion of tissue in an iodine solution to rid the embryos of any amebas and micrometazoa. When culturing whole colonies of Botryllus schlosseri at the University of Washington, Tacoma, we find many microscopic organisms inhabiting the tissue and tunic of our field collected colonies. Currently, we are unable to successfully rear colonies in our recirculating seawater system and so we are considering extracting tissue from field colonies. In order to do so we need to be able to eliminate the microfauna parasitizing our colonies and I believe dipping those colonies briefly in an iodine solution may aid in this process.