A human diploid fibroblast cell line has been established from the lung tissue of a male fetus. This has been characterized and frozen away in large quantity. A smaller quantity of fibroblastlike cells from skin has also been established, partially characterized, and placed in frozen storage from the same fetus. This project is in support of the National Institute on Aging research in general cell biology. The present lines designated IMR-91 lung and IMR-91 skin complement the previous human diploid fibroblast culture (IMR-90) established from a female fetus. The lack of random inactivation of one of the two X chromosomes in the present male line reduces the genetic heterogeneity inherent in the female line.

The cooling of a heated electron gas by electron impact excitation of CO2 is believed to be an important process in the neutral atmospheres of Mars and Venus. Electron energy loss rates are calculated for a variety of excitation processes as a function of electron temperature at a CO2 gas temperature of 200 K. Vibrational excitation is found to be the most efficient cooling mechanism for the range of electron temperatures studied. The contributions of various vibrational energy loss processes to electron cooling are discussed.

A new human diploid fibroblast-like cell line has been established from lung tissue of a female fetus. This has been frozen away in large quantity and characterized for use in research and related purposes. This is the first of a planned series of human cell lines to be established, characterized, and banked in large quantity in support of the National Institute on Aging research and general cell biology.

This chapter focuses on human biopsy material from genetic abnormalities. Among the most useful tissue cultures available to the geneticist are those derived from human skin. Skin biopsies are easily obtained, the cultures derived from them are easy to grow, they can be frozen for years in liquid nitrogen, and recovered in a highly viable state. The skin specimen may be collected by simple punch biopsy. In the method described in the chapter, stainless steel punches together with forceps and scissors are cleaned, wrapped, and autoclaved. The skin is swabbed with alcohol, rinsed with sterile saline, and dried with sterile gauze. Almost any area of the body can be used including the buttocks, the back medial to the upper border of the scapula, the flexor surface of the forearm, and the deltoid region. The procedure is simple, rapid, and an anesthetic is usually not required. If it is considered essential to use one, an intradermal injection of several milliliters of 285 procaine solution will anesthetize the skin.

Forty-six cell cultures established from amniocentesis fluids were preserved in liquid nitrogen and later recovered from the frozen state with little loss of viability as compared to prefreeze viability. Five to 10% glycerol was found to be optimal for preservation in liquid nitrogen, and as few as 5×105 viable cells per frozen ampule could initiate cell growth. Storage in liquid nitrogen did not affect the genetic stability of glucose-6-phosphate dehydrogenase, lactate dehydrogenase, malic dehydrogenase, leucine aminopeptidase, acid phosphatase, or 6-phosphogluconic acid dehydrogenase isozymes of the amnion cultures.

descriptions of cell cultures from individuals with chromosome aberrations.

Three mosquito cell cultures designated as Suitor's clone ofAedes aegypti, Culiseta inornata andAedes vexans were shown to be moth by immunological, karyological, and isozyme analyses. The cells reacted with rabbit antimoth serum but not rabbit antimosquito serum. Chromosome analyses indicated Lepidopteran rather than Dipteran morphology, and three isozyme systems were confirmative. Any one of these assays would be sufficient to indicate that contamination had occurred and could be used as a periodic check for identity of cell cultures. Morphology and growth characteristics are also valid criteria to distinguish between these particular orders of insect cells.

During the past two decades we have witnessed an increasing accumulation of biological knowledge in cytogenetics, biochemistry, immunology and biology related to experimental techniques developed in mammalian cell cultures. Comparable achievements are being reported from research studies in poikilothermic cell cultures. It is now obvious that insect cell cultures pioneered by many of the authors in this volume and by the late Dr. Suitor will also allow us to gain further insight into many biological phenomena associated with insects.

For a number of years this laboratory has been one of those engaged in a program to develop a Central Cell Line Bank and Registry for the characterization and preservation of valuable cell cultures (1).

Fresh human skin placed in plasma clot culture shows outgrowth of epithelial cells within 3 days followed 3 to 6 days later by fibroblasts. Plasma clot culture of human skin pretreated in medium containing 10% glycerol at 4°C, frozen, and thawed showed outgrowth of fibroblasts only. Using this increased susceptibility of epithelial cells to freeze injury as a criterion, we developed a freezing method in which skin is pretreated with 20 to 30% glycerol at 4°C for 1 to 2 hrs before freezing. Human skin preserved in liquid nitrogen by this method grew out in plasma clot culture very much like fresh skin.

Tissue culture media of various types have been devised in the past few years to support the growth of living cells for many generations vitro . We have made modifications in one tissue culture medium (Eagle's minimum essential medium, MEM) so that it will support the functional viability of rabbit hearts stored for 48 hrs at 4°C. Rabbit hearts perfused with Eagle's MEM with added KCl to give a final concentration of 13 to 15 mEq per liter of K + stop contracting within 1 min. Perfusion with the high K + medium is then continued at 4°C. After a series of 40 experiments it has been established that rabbit hearts perfused with Eagle's MEM containing 143 mEq of Na + per liter, 13 to 15 mEq of K + per liter, 9.8 mEq of Ca ++ per liter, and 0.2% glucose equilibrated with 95% O 2 -5% CO 2 can be preserved at 4°C for 24 to 48 hrs with 100% recovery of function as measured by the force of contraction.

MACCLEAN1,2 has indicated that magnetic fields were effective in arresting carcinoma growth in patients, and Barnothy3 stated that magnetic fields, in some way, retarded the ageing (hence growth) process in mice.

In view of the reported manifestation of latent tetanus infections in humans exposed to stress or shock, this subject was investigated experimentall by using mice. Latent tetanus infection was roduced by the injection of washed tetanus spores under aseptic conditions. Standard shock was produced by the Noble‐Collip method; standard degree of exhaustion was produced by employing the swim technique and by exposing the mice to low temperawes. Manifestation of the disease was not evidenced in any of these instances. On the other hand, active tetanus was induced by the subsequent injection of calcium chloride solutions. The relationship between shock, stress, and tetanus infection is discussed.

Although it had been reported that the antihistamine Phenergan increases the spread of bacterial and viral infections in the host, such an effect was not observed in tetanus infection after the administration of Phenergan. Antihistamines such as Phenergan appear not to produce any marked increase in tetanus fatalities or symptoms, even though they cause spreading of the microorganisms.