Biochemical and metabolic properties of bovine type II pneumocytes in primary culture

Lung ◽  
1989 ◽  
Vol 167 (1) ◽  
pp. 343-350 ◽  
Author(s):  
H. G. Augustin-Voss ◽  
H. A. Schoon ◽  
A. Gerull ◽  
S. Ueberschär
Keyword(s):  
Primary Culture ◽  
Type Ii ◽  
Type Ii Pneumocytes ◽  
Metabolic Properties ◽  
Bovine Type

The effect of stone-wool on rat lungs and on the primary culture of rat alveolar macrophages and type II pneumocytes

2005 ◽  
Vol 26 (1) ◽  
pp. 16-24 ◽  
Author(s):  
Erzsébet Tátrai ◽  
Márta Brozik ◽  
Ágnes Drahos ◽  
Zuzana Kováčiková ◽  
Éva Six ◽  
...  
Keyword(s):  
Primary Culture ◽  
Alveolar Macrophages ◽  
Type Ii ◽  
Rat Lungs ◽  
Type Ii Pneumocytes ◽  
Stone Wool

scholarly journals Effects of Xanthine Derivatives on Phosphatidylcholine Secretion in Primary Culture of Rat Type II Pneumocytes

1995 ◽  
Vol 67 (2) ◽  
pp. 165-168 ◽  
Author(s):  
Manabu Okumura ◽  
Michio Tsuruoka ◽  
Yoichiro Isohama ◽  
Hirofumi Kai ◽  
Kazuo Takahama ◽  
...  
Keyword(s):  
Primary Culture ◽  
Type Ii ◽  
Xanthine Derivatives ◽  
Type Ii Pneumocytes ◽  
Phosphatidylcholine Secretion

Density-independent isolation of type II pneumocytes after partial pneumonectomy

1989 ◽  
Vol 256 (3) ◽  
pp. C515-C521 ◽  
Author(s):  
B. D. Uhal ◽  
G. D. Hess ◽  
D. E. Rannels
Keyword(s):  
Primary Culture ◽  
Compensatory Growth ◽  
Thymidine Incorporation ◽  
Maximal Velocity ◽  
Type Ii ◽  
Percoll Gradient ◽  
Type Ii Pneumocytes ◽  
The Right ◽  
Immunoglobin G ◽  
Exogenous Spermidine

Type II pneumocytes were isolated by immunoglobin G (IgG) panning from the lungs of normal rats and the right lung of rats (180-200 g) subjected to left pneumonectomy. Cells were studied at 7 (pnx-7) and 15 (pnx-15) days postoperative, times during and after, respectively, rapid compensatory growth of the right lung. After 24 h of primary culture, pnx-7 cells contained 32% more protein per DNA, and incorporated thymidine at a rate 224% greater than cells isolated from control rats. Both the protein-to-DNA ratio and thymidine incorporation returned to control values in pnx-15 cells. Uptake of exogenous spermidine also was increased by 50% in pnx-7 cells at 24 h of primary culture and returned to control values in pnx-15 cells. Increased spermidine uptake was due to an increase in the maximal velocity (Vmax) of transport from 30.3 (control) to 45.5 pmol.micrograms DNA-1.h-1 (pnx-7), with no change in the apparent Km of 1.32 microM. No change was observed in the relative rates of phospholipid or neutral lipid biosynthesis. The increases in thymidine incorporation and spermidine uptake were significantly greater than those previously observed [Am. J. Physiol. 254 (Cell Physiol. 23): C684-C690, 1988] in pnx-7 cells isolated by Percoll gradient sedimentation. These results suggest that pnx-7 lungs contain distinct subpopulations of type II pneumocytes, the recovery of which is dependent on the cell isolation protocol employed.

Alterations in type II pneumocytes cultured after partial pneumonectomy

1988 ◽  
Vol 254 (5) ◽  
pp. C684-C690 ◽  
Author(s):  
S. R. Rannels ◽  
D. E. Rannels
Keyword(s):  
Primary Culture ◽  
Calf Serum ◽  
Serum Levels ◽  
Type Ii ◽  
Rapid Phase ◽  
Culture Time ◽  
Pulmonary Epithelial Cells ◽  
Type Ii Pneumocytes ◽  
The Right

Type II pulmonary epithelial cells prepared from the lungs of normal rats were compared in primary culture to cells derived from the right lung of animals subjected previously to left pneumonectomy (PNX). Studies were initiated on the sixth post-PNX day, during the rapid phase of compensatory right lung growth. After 24 h in vitro, PNX cells were 30-40% larger than controls and contained 20-50% more DNA. The magnitude of these differences was dependent on serum concentration (fetal calf serum; 1 and 10%, respectively) and, under most conditions, decreased as culture time was extended to 48 or 72 h. Incorporation of [3H]thymidine into DNA was also elevated (greater than 50%) on the first culture day in the PNX group at both serum levels, and remained so through day 3 at low serum, as thymidine incorporation became more rapid in all cells. Similarly, rates of spermidine uptake were elevated in cells prepared from lungs of PNX animals on culture day 1, but this effect too was lost by day 3. Thus type II pneumocytes isolated from the lungs of PNX rats exhibit metabolic changes typical of accelerated cell growth at early intervals of primary culture in vitro. Although these changes are lost as culture time is extended and the cells lose differentiated characteristics, the results suggest that such pneumocytes may provide useful information regarding factors which regulate compensatory growth of lung tissue.

Isolation of bovine type II pneumocytes in high yield and purity

Lung ◽  
1989 ◽  
Vol 167 (1) ◽  
pp. 1-10 ◽  
Author(s):  
H. G. Augustin-Voss ◽  
H. A. Schoon ◽  
N. Stockhofe ◽  
S. Ueberschär
Keyword(s):  
High Yield ◽  
Type Ii ◽  
Type Ii Pneumocytes ◽  
Yield And Purity ◽  
Bovine Type

Increased surfactant internalization by rat type II cells cultured on microporous membranes

1993 ◽  
Vol 264 (3) ◽  
pp. L300-L307 ◽  
Author(s):  
M. R. Chinoy ◽  
C. Dodia ◽  
A. B. Fisher
Keyword(s):  
Primary Culture ◽  
Lung Surfactant ◽  
Light And Electron Microscopy ◽  
Cell Protein ◽  
Type Ii Cells ◽  
Type Ii ◽  
Natural Surfactant ◽  
Culture Cells ◽  
Type Ii Pneumocytes ◽  
Cells Cultured

We evaluated the influence of various substrata in maintaining internalization of biosynthesized natural surfactant by type II pneumocytes in primary culture. Cells were incubated for 2 h with 3H, 35S-labeled surfactant, prepared by perfusing isolated rat lungs with [3H]-choline + [35S]methionine, and analyzed for trypsin-resistant radiolabel. At 24 h of culture, uptake of [3H]phosphatidylcholine (PC) was 71.9 +/- 6.1 micrograms/mg protein and 35S-protein was 12.4 +/- 2.4 micrograms/mg cell protein for cells cultured on Transwell membranes. Uptake with this substratum was significantly greater than for cells cultured on plastic, Primaria, or Transwell-COL and was maintained for 72 h of culture. [3H]PC/35S-protein (micrograms/micrograms) for cells on Transwell was 5.8 +/- 0.7 compared with 3.4 +/- 0.1 in isolated surfactant. Results were similar at 72 h of culture. 8-Bromoadenosine 3',5'-cyclic monophosphate or terbutaline significantly stimulated the uptake of both surfactant components for cells on Transwell, but not by cells cultured on plastic. By light and electron microscopy, cells cultured on Transwell maintained characteristics of granular pneumocytes, including cuboidal shape, lamellar bodies, and microvilli. We conclude that the use of Transwell microporous membrane for primary culture of rat type II cells maintains their differentiated characteristics as assessed by the uptake of lung surfactant and its response to beta-adrenergic agonists.

Culture of type II pneumocytes on a type II cell-derived fibronectin-rich matrix

1987 ◽  
Vol 253 (6) ◽  
pp. C759-C765 ◽  
Author(s):  
S. R. Rannels ◽  
C. S. Fisher ◽  
L. J. Heuser ◽  
D. E. Rannels
Keyword(s):  
Extracellular Matrix ◽  
Dna Synthesis ◽  
Primary Culture ◽  
Type Ii Cells ◽  
Type Ii ◽  
Type Ii Pneumocytes ◽  
Plastic Surface ◽  
Type Ii Cell ◽  
Differentiation And Proliferation

Recent evidence suggests that during primary culture, type II pneumocytes synthesize and deposit components of an extracellular matrix. The present study investigated the response of freshly isolated type II cells to a preformed, fibronectin-rich matrix synthesized by type II cells over a 6-day interval of primary culture on a plastic surface. Type II cells on 6-day matrix (M6) degraded the preformed matrix and deposited newly synthesized fibronectin more rapidly than cells on plastic, suggesting that M6 itself stimulated type II cell-mediated matrix turnover. In type II cells on plastic, incorporation of radiolabeled thymidine into DNA increased 620 and 1,880% after 2 and 3 days in culture, respectively, as the cells assumed a more flattened phenotype. Although cells on M6 did not divide, both basal rates of thymidine labeling and sensitivity to serum modulators of DNA synthesis were enhanced by the M6 surface, as compared with plastic. Culture of type II cells on surfaces of purified fibronectin enhanced the rate of DNA synthesis in a manner similar to that observed on M6; this effect was blocked by antifibronectin. The data suggest that more rapid fibronectin synthesis and deposition are important components of the response of type II cells to primary culture. Extracellular matrix produced by type II cells appears to be similar to the basement membrane onto which these cells proliferate in vivo after lung injury. A fibronectin-rich surface in itself may thus induce additional extracellular matrix synthesis and further direct cellular differentiation and proliferation.

The Identification of Phospholipid Micelles in Glutaraldehyde-Urea Embedded Tissue

1975 ◽  
Vol 33 ◽  
pp. 494-495
Author(s):  
Daniel C. Pease
Keyword(s):  
Electron Micrographs ◽  
Lung Tissue ◽  
Osmium Tetroxide ◽  
Uranyl Acetate ◽  
Type Ii ◽  
Lead Citrate ◽  
Phospholipid Micelles ◽  
Type Ii Pneumocytes ◽  
Ultrathin Sections ◽  
Polar Water

It is reasonable to think that phospholipid micelles should be visible and identifiable in electron micrographs of ultrathin sections if only they can be preserved throughout the embedding process. The development of highly polar, water-containing, aminoplastic embedments has made this a likely possibility. With this in mind, an investigation of the lecithin-secreting, Type II pneumocytes of the lung is underway.Initially it has been easiest to recognize phospholipid micelles in lung tissue fixed first with glutaraldehyde, and then secondarily exposed to osmium tetroxide. However, the latter is not a necessary concomitant for micellar preservation. Conventional uranyl acetate and lead citrate staining is finally applied. Importantly, though, the micelles have been most easily seen in tissue embedded in 507. glutaraldehyde polymerized with urea, as described in detail by D.C. Pease and R.G. Peterson (J. Ultra- struct. Res., 41, 133, 1972). When oriented appropriately, the micellar units are seen as tiny, bilayer plates.

Sign in / Sign up

Export Citation Format

Share Document