ANALYSIS OF THE GROWTH KINETICS OF A HUMAN LYMPHOMA CELL LINE

The growth kinetics of an established human lymphoma cell line were analyzed by a variety of techniques utilizing various cell inocula (5 x 10⁴ - 5 x 10⁵ cells) dispensed into 60 mm diameter dishes. Techniques included pulse-labeled mitosis (PLM), continuous labeling with ³H-TdR, time-lapse photography (TLP), cell counts by electronic particle counter, and DNA histography obtained by pulse cytophotometry (PCP). There were no significant differences among values determined for any kinetic parameters as a function of cell concentration. the average doubling time of exponentially growing cells, regardless of cell inoculum, was 44.1 hr. the generation time determined by PLM was 31.1 hr with a SD of 4.7 hr. Transit times for each stage were: T_G1= 10.6 hr, T_s= 9.9 hr, T_G2= 9.9 hr, and T_m= 0.7 hr. Repeated experiments using continuous labeling with ³H-TdR demonstrated a T_G2 of 6.3 hr. the longer value determined by PLM is possibly due to the technical manipulations of this procedure which may delay pulse-labeled cells from resuming cell cycle transit. Hence, values for cell cycle stages were recalculated to give T_G1= 14.1 hr, T_s= 9.9 hr, T_G2 = 6.3 hr, and T_m= 0.7 hr. These results were used to compute the size of each cell cycle stage compartment pool and corresponded very closely to values defined directly by PCP. TLP analysis considered only cells that produced colonies of at least thirty-two cells. Generation times ranged from 8 to 89 hr and showed a positive skewness. the average value measured for 330 divisions was 34.5 hr with a SD of 13.2 hr. Thus, the variance predicted by curve fitting of the PLM data did not correlate with that defined by time-lapse photography nor did it encompass the range in generation times observed directly by TLP. There was a positive correlation between sister-sister cell generation times (+0.66) but no relation was noted for mother-daughter values.     

GROWTH AND CELL POPULATION KINETICS OF SINGLE AND MULTIPLE KHT SARCOMAS

KHT sarcomas were implanted in syngeneic C3H/Km mice either as single intradermal tumors on the flank or as nine or ten intradermal tumors on the back. The growth curves of the single and multiple tumors were indistinguishable throughout the accurately measurable volume range. The cell population kinetics of the single and multiple tumors, examined by the per cent labeled mitoses technique, were not significantly different in any respect. The cell population kinetics in the center and periphery of the single tumors were examined separately and found to be the same.     

CELL KINETICS OF A CHONDROSARCOMA

The cell kinetics of the transplantable DC-II mouse chondrosarcoma have been studied by the pulse labelled mitoses method. The analysis gave the following estimates for the phases of the cell cycle: G, 10-5 hr; S, 9-5 hr; G₂, 4 hr with an intermitotic time of 23-5 hr. Consideration of the overall growth of the tumour indicated that the growth fraction and cell loss factor both had values of about 0–5. The results are compared with cell kinetic data from sarcomas and other cartilage tissues.     

CELL KINETICS OF GROWTH CARTILAGE IN THE RAT TIBIA II. MEASUREMENTS DURING AGEING

A number of cell kinetic techniques using labelled thymidine and autoradiography have been applied to study growth cartilage in the rat tibia during ageing. No change in the duration of the synthesis phase was found from 4 to 13 weeks of age but there was a reduction in cell proliferation rate during this period. Measurements of labelling index, proliferation zone size and height of hypertrophic cells were used to calculate the growth rate of the bone from 7 days to 1 year. The results agreed well with radiographic measurements of bone growth.     

CELL KINETICS OF GROWTH CARTILAGE IN THE RAT TIBIA I. MEASUREMENTS IN YOUNG MALE RATS

Cell kinetic parameters for the proximal growth plate of the tibia have been measured in young rats. Analysis of a pulse labelled mitosis study gave values of 55 ± 40 hr for the cycle time and 6.5 ± 0.3 hr for the synthesis time in 6-week-old rats. The results of a simulated continuous labelling experiment agreed with this data and provided further information on the size and proliferation rate of the stem cell zone. Diurnal variations in mitotic index and labelling index in the tissue have been investigated.     

THE CELL POPULATION KINETICS OF NEUTROPHILIC CELLS

Autoradiographic data for the entry of tritiated thymidine labelled cells into the post-proliferative neutrophilic cell compartments following a single injection of isotope have been analysed in terms of two cell kinetic models which differ in the assumed relationships between cell maturation and division. Comparisons with the experimental data were made in an attempt to assess the validities of the models, and kinetic parameters for the compartments of recognizable neutrophilic cells were estimated. Control mechanisms which have been proposed for the granulocyte system are discussed in terms of the kinetic models which were chosen in their determination. Although it was not possible to make a clear choice between the proposed models, preference was established for a random model which did not involve cell loss.     

MATRIX SIMULATION OF DUODENAL CRYPT CELL KINETICS

Steady state crypt cell kinetics have been simulated using matrix algebra. The model crypt cell population is distributed through two proliferation compartments (P₁ and P₂) and a quiescent state (Q). Under steady state conditions half the daughter cells produced on completion of P₁ enter G₁ of P₂ and half enter G₁ of P₁. Both P₂ daughter cells enter Q. Cells in Q are non-dividing but retain the potential to divide. On completion of Q, cells lose the potential to divide and move up onto the villi. The model has been developed by simultaneously simulating the following biological data: (1) the per cent labeled mitosis (PML) curve, (2) the number of labeled cells per crypt as a function of time following an injection of ³H-thymidine, and (3) the total number of cells per crypt.     

CELL POPULATION KINETICS OF EXCISED ROOTS OF PISUM SATIVUM

The cell population kinetics of excised, cultured pea roots was studied with the use of tritiated thymidine and colchicine to determine (1) the influence of excision, (2) the influence of sucrose concentration, (3) the average mitotic cycle duration, and (4) the duration of mitosis and the G₁, S, and G₂ periods of interphase.¹ The results indicate that the process of excision causes a drop in the frequency of mitotic figures when performed either at the beginning of the culture period or after 100 hours in culture. This initial decrease in frequency of cell division is independent of sucrose concentration, but the subsequent rise in frequency of division, after 12 hours in culture, is dependent upon sucrose concentration. Two per cent sucrose maintains the shortest mitotic cycle duration. The use of colchicine indicated an average cycle duration of 20 hours, whereas the use of tritiated thymidine produced an average cycle duration of 17 hours.     

CELL KINETICS IN THE SEBACEOUS GLANDS OF THE MOUSE. II. THE GLANDS DURING HAIR GROWTH

The sebaceous glands of the mouse have been studied during hair growth initiated either spontaneously or artificially. The labelling index of the glands increases early in the spontaneous hair growth period. That of the epidermis is much lower and hardly changes during the growth period. After the initiation of hair growth by plucking, changes in cell proliferation in the sebaceous glands appear to follow those in the epidermis. The size of the gland and the number of cells in it also change after plucking. These variations can be related to the stages of hair growth.     

RADIATION EFFECTS ON THE GROWTH RATE AND CELL POPULATION KINETICS OF ACTIVELY GROWING AND DORMANT ROOTS OF TRADESCANTIA PALUDOSA

Actively growing and dormant roots of Tradescantia paludosa were exposed to x-rays to compare the radiosensitivity of an actively proliferating tissue with that of one which is not active but is potentially proliferative. The level of effect was ascertained by the degree of change in the rate of root growth 4 days after exposure. Cell population kinetics were measured in control and in irradiated roots to determine whether or not a change was produced either in the number of proliferating cells or in the mitotic cycle duration which was sufficient to explain the altered rate of root growth. Nuclear volumes were also measured to provide an estimate of the relative total target size in actively growing vs. dormant roots. Tritiated thymidine was used to measure the cycle duration and the proportion of cells synthesizing DNA. The results showed that 184 and 305 r respectively were required to reduce the linear root growth rate to 37 per cent of that of the control for actively growing and dormant roots. Mitotic cycle duration, measured 4 days after x-ray exposure, was the same as in the control. The number of proliferating cells, however, was reduced. The rate of cell production in the irradiated roots was reduced to approximately one-half that of the controls. The average nuclear volumes of active and dormant roots were 733 and 491 µ³ respectively; thus the difference in the number of roentgens required to reduce growth to 37 per cent of that of the control can be attributed to the different average nuclear volumes. Therefore, the experiments suggest that part if not most of the differences in sensitivity between an actively dividing and an essentially non-dividing meristematic cell population resides in their different average nuclear volumes. Thus the law of Bergonie and Tribondeau needs to be reinterpreted, since the basic reason for the differences is secondary to whether or not the meristematic cells are proliferating.     

细胞增长的数学模型

本文研究一个关于细胞增长的数学模型,用微分方程组来确定。通过系统的稳定性的分析指出参数对细胞组织容量的平衡状态的影响。     

KINETICS OF GROWTH OF HAEMOPOIETIC COLONY CELLS IN AGAR

Cell kinetic parameters of mouse granulocytic and mononuclear cells growing in colonies in agar cultures have been measured. Analysis of flash and continuous labelling studies with ³H-thymidine together with determinations of colony size, growth fraction and mitotic indices, gave the following values for the phases of the cell cycle: G₁= 6·3 1·6 hr, S = 5·8 ± 1·4 hr, G₂= 1·7 ± 0·1 hr and M = 0·7 ± 0·1 hr (42 ± 8 min). No difference in the cell cycle parameters of granulocytic and mononuclear cells were found in this study.
Colonies of different size from cultures of the same age group had similar labelling indices, indicating that the size of a colony is not a function of the rate of proliferation of cells in the colony. Rather, variation in colony size is probably representative of an initial delay in the onset of colony development.     

双歧杆菌生长和代谢过程的研究

研究了双歧杆菌在两种培养基（牛乳培养基和肉汤培养基）中的生长和代谢的规律,测定了厌氧发酵过程中菌体生长及基质消耗曲线,探索了在发酵过程中流加碱调节ｐＨ值以提高产菌量的途径。     

KINETICS OF MURINE HAEMOPOIETIC CELL PROLIFERATION IN DIFFUSION CHAMBERS

Murine bone marrow cells were cultured in cell impermeable diffusion chambers in the abdominal cavities of mice. The kinetics of granulocyte and macrophage formation were studied by stathmokinetic and autoradiographic techniques. During the period of most rapid growth of proliferative granulocytes, their generation time and its different phases were: t _c∽ 8 hr, t _G1∽ 1·5 hr, t _s∽ 5·5 hr, t _G2∽ 0·7 hr and t _M∽ 0·25 hr.
The generation time of macrophages and their precursors was approximately 8 hr. Formation of macrophages was significantly reduced when chamber inoculum was increased, as judged by ³H-TdR labelling index.     

KINETICS OF CELL RENEWAL, CELL MIGRATION AND CELL LOSS IN THE HAIRLESS MOUSE DORSAL EPIDERMIS

Forty hairless mice were given injections of tritiated thymidine every 4th hour during 10 days. At 24 hr intervals groups of four mice were killed. The numbers of labelled basal and differentiating cells were determined by autoradiography with a stripping film technique. To determine the background activity skin sections from uninjected control mice were subjected to the same stripping film procedure. Another group of hairless mice was given one single pulse labelling with tritiated thymidine. The number of labelled mitoses was scored for 12 hr after the injection. At 10, 12 and 15 hr after the injection, the numbers of labelled basal and differentiating cells were also determined. A mathematical model of cell population kinetics in the epidermis has been suggested. The results of different simulations on this model were compared with the observed results. The curve of mean grain counts under continuous labelling increased from day to day with two well-defined plateaux. The percentage of all labelled cells increased rapidly up to the 3rd day, and thereafter the curves gradually flattened off. When basal cells and differentiated cells were considered separately the labelling index of the basal cells increased rapidly for the first 3 days and then flattened off at the 100% level on the 5th day. The labelling index of the differentiating cells was low during the first 3–4 days. Then a steep increase in the percentage of labelled differentiating cells was seen, but the curve flattened off again close to the 100 % level after the 7th day. The labelled mitosis curve had its maximum 5 hr after the thymidine injection. The curve fell again to almost zero at 12 hr. Ten, 12 and 15 hr after the injection, 6, 7 and 7% respectively of the labelled cells were found in the spinous layer. It was concluded that three grains over each nucleus could be used as lower limit for considering a cell as labelled. On this basis, tritiated thymidine injections every 4th hour can be considered as continuous labelling.