A.Fi and D-1 Re Cell Lines Overshadowed By HeLa Cell Line

While reading chapter 13 of Rebecca Skloot's "Immortal Life of Henrietta Lacks," which discusses the sudden progress and growth of the field of cell/tissue culture after Henrietta Lacks' death; I happen to come across one intriguing paragraph. The paragraph states "... there were other cells to work with, including some he'd grown himself: A.Fi and D-1 Re... but they were harder to culture, so they never took off like Henrietta's cells" (Skloot, 2010, p.103). I found this very intriguing because I did not know that other continuous cell lines were grown by George Guy aside from HeLa Cells. I began to think that if these cells were just as efficient and as easy to culture as HeLa then the rate of progress in the field of Cell/Tissue culture as well as other fields in science would have been much faster and greater technological advances in fields such as molecular biology, genetics, virology, physiology, immunology, as well as biotechnology and pharmaceutical industries, etc. would have been achieved sooner. I began to wonder, what made the A.Fi and D-1 Re harder to culture, when compared to the HeLa cell line, and how much they deviate to the conditions required for HeLa cell lines to grow (physiochemical environment, physiological conditions, microenvironment).

In order to determine the properties of both A.Fi and D-1 Re and the conditions they are required for them to be cultured, I started by searching for articles using various databases and low and behold I found Dr. George Gey's original November 1954 article from the Annals of the New York Academy of Sciences, titled "ACTIVITIES AND RESPONSES OF LIVING CELLS AND THEIR COMPONENTS AS RECORDED BY CINEPHASE MICROSCOPYAND ELECTRON MICROSCOPY." (Gey, Shapras, Borysko, 1954). The main objective of this article was to utilize a new technique (at the time) called cinephase microscopy to various cell cultures in order to monitor the growth, proliferation, and differentiation as well as make comparisons of the cell's composition and structure of various cell lines by. They were able to utilize:

A series of static images and short motion picture scenes were used to illustrate some of the similarities and differences in the structural organization and the behavior of various components of the

tumorous cell strain of the rat (T-333) derived in vitro from a normal fibroblast strain 1 4 ~ .C~ine phase studies were also made of the 23-year-old strain of human chondromyxosarcoma, strain D-1 Re; of the 15-year-old human fibrosarcoma, strain A.Fi.; and of the 3-year-old strain of human epidermoid carcinoma of the cervix, strain HeLa, when grown in thin tissue-culture slides (Gey, Shapras, Borysko, 1954).

This study allows one to actually visualize the physical difference between HeLa cells as well as the A.Fi., and D-1 Re cells, as well as determine the reason why they are difficult to culture compared to HeLa cells.

They monitored the development of pseudopodia in all cells as well as their increase in size and morphology using cinephase microscopy. When the T-333 strain was compared to the wild type rat cells once can see

that T-333 was:

(1) thicker; (2) to show a prominent ring of inclusion droplets distributed around the nucleus and juxtanuclear area; (3) to have a large juxtanuclear area so ridged as to bend the large polyploid nucleus found in many malignant cells...(4) to manifest increased feeding habits through abundant pinocytosis; and (5) to have a larger centrosphere in

the region shown by the arrow. Also, the tumor-cell derivatives show less cohesion of the cells of their colonies and fewer microfibrils (Gey, Shapras, Borysko, 1954).

The human cell lines HeLa, A.Fi., and D-1 Re cells displayed similar morphology as the T-333 cells and most importantly, mitochondrial distortion and size reduction is present in all tumorous cell lines, which can be used to determine the efficiency and ease of culture. One can see that in HeLa:

Within an

eight-minute period, inclusion droplet No. 1 engages mitochondrion No. 2, and su

cceeds in breaking it within three

minutes... At this time, it has moved a considerable

distance

and has now engaged a second mitochondrion, No. 3, and within

two minutes it stretches mitochondrion No. 3 to form two

broken fragments At the six-minute interval... it has already engaged a third mitochondrion No. 4 and may be seen to move still farther on its way toward the juxtanuclear mass. These events reveal the dramatic movement of such components within 10 minutes (Gey, Shapras, Borysko, 1954).

One can see on the A.Fi and the D-1 Re cells that the movement of components are slower which may indicate slower growth which is more difficult to culture. However, there are still other factors that are not elaborated upon in this study which may greatly contribute to the A.Fi an

D-1 Re's inability to be cultured at the level of HeLa. Even though I have not reached a definite conclusion, I was able to see what these cells look like, and it gave me an idea that it is very possible for other cells to be immortalized and used for cell cultures.

...........HeLa cells.................D-1 Re cells...............A.Fi cells