DIVISION OF BALB-C MOUSE 3T3 AND SIMIAN VIRUS-40-TRANSFORMED 3T3 CELLS IN CELLULAR AGGREGATES - (DNA SYNTHESIS DENSITY-DEPENDENT INHIBITION OF PROLIFERATION ANCHORAGE DEPENDENCE OF GROWTH SERUM STIMULATION
DIVISION OF BALB-C MOUSE 3T3 AND SIMIAN VIRUS-40-TRANSFORMED 3T3 CELLS IN CELLULAR AGGREGATES - (DNA SYNTHESIS DENSITY-DEPENDENT INHIBITION OF PROLIFERATION ANCHORAGE DEPENDENCE OF GROWTH SERUM STIMULATION
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DOI：
10.1073/pnas.74.9.3874
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发表时间：
1977-01-01
影响因子：
11.1
通讯作者：
GERSHMAN, H
中科院分区：
文献类型：
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作者：
CARRINO, D;GERSHMAN, H
将BALB/c小鼠3 T3成纤维细胞和由SV 40转化的3 T3细胞在搅拌的液体培养基中培养为聚集体。当维持每天更换培养基时，3 T3细胞将[3 H]胸苷掺入酸不溶性物质中的速率为3 T3细胞在静止期平板培养物中的1/4，但为相同细胞在静止期平板培养物中的16倍。SV 40-转化的3 T3细胞聚集体掺入[3 H]胸苷的速率是静止期平板培养中SV 40 - 3 T3细胞的1/3，是静止期平板培养中这些细胞的14倍。在[3 H]尿苷和[3 H]胸苷存在下孵育2小时的3 T3和SV 40 - 3 T3细胞聚集体的放射自显影表明，在此期间核苷渗透到聚集体中仅限于外部4-6细胞层。还在存在放射性标记的胸苷和尿苷的情况下连续孵育聚集体4天。在这些条件下，核苷渗透不受限制，100%的SV 40 - 3 T3细胞和56%的3 T3细胞将[3 H]胸苷掺入酸不溶性物质中。通过不受运输、合并液大小或渗透可能变化影响的技术，测定3 T3和SV 40 - 3 T3细胞聚集体中的细胞分裂、细胞损失和净细胞蓄积率。SV 40 - 3 T3细胞分裂，总群体的倍增时间为26.0 h。由于细胞损失，总细胞数增加得更慢（倍增时间为48.3 h），其发生的半衰期（50%细胞损失的时间）为53.3 h。聚集体中的3 T3细胞仅在3天后开始分裂，然后以76.4 h的总群体倍增时间进行分裂。总细胞数减少（半衰期为26.3 h），因为细胞分裂速率超过细胞损失速率，细胞损失速率恒定，半衰期为22.9 h。结果表明，3 T3和SV 40 - 3 T3细胞在聚集体中显示出与它们先前在常规平板培养中报道的行为一致的生长特性：SV 40转化的3 T3细胞可以在高细胞密度条件下增殖到比3 T3细胞大得多的程度。这两种细胞系显示出相对于融合平板培养物的聚集体分裂能力增加，尽管细胞密度高且不存在对人工固体基质的锚定。
BALB/c mouse 3T3 fibroblast cells and 3T3 cells transformed by SV40 were cultured as aggregates in agitated liquid medium. When maintained with daily medium changes, 3T3 cells incorporated [3H]thymidine into acid-insoluble material at a rate 1/4 that of 3T3 cells in logarithmic-phase flat cultures but 16 times that of the same cells in stationary-phase flat cultures. SV40-transformed 3T3 cells in aggregates incorporated [3H]thymidine at 1/3 the rate of SV40-3T3 cells in logarithmic-phase flat culture and 14 times that of these cells in stationary-phase flat culture. Autoradiographs of aggregates of 3T3 and SV40-3T3 cells incubated for 2 h in the presence of [3H]uridine and [3H]thymidine indicated that penetration of the nucleosides into aggregates during this period was limited to the outer 4-6 cell layers. Aggregates were also incubated in the presence of radiolabeled thymidine and uridine continuously for 4 days. Under these conditions, where nucleoside penetration was not limiting, 100% of the SV40-3T3 cells and 56% of the 3T3 cells incorporated [3H]thymidine into acid-insoluble material. The rates of cell division, cell loss and net cell accumulation in aggregates of 3T3 and SV40-3T3 cells were measured by techniques not influenced by possible alterations in transport, pool size or penetration. SV40-3T3 cells divided with a doubling time for the total population of 26.0 h. The total cell number increased more slowly (doubling time of 48.3 h) because of cell loss, which occurred with a half-time (time for 50% of the cells to be lost) of 53.3 h. The 3T3 cells in aggregates began to divide only after 3 days, then did so with a doubling time for the total population of 76.4 h. Total cell number decreased (half-time of 26.3 h) because this rate of cell division was exceeded by the rate of cell loss, which was constant with a half-time of 22.9 h. The results suggest that 3T3 and SV40-3T3 cells display growth properties in aggregates consistent with their previously reported behavior in conventional flat culture: SV40-transformed 3T3 cells can proliferate under conditions of high cell density to a much greater extent than 3T3 cells. Both cell lines display an increased capacity to divide in aggregates relative to confluent flat culture, despite conditions of high cell density and absence of anchorage to an artificial solid substrate.