Recovery of rat type II cell surfactant components during primary cell culture.
A culture system designed to maintain the differentiated characteristics of rat type II cells based on protocols used for human fetal lung pneumocytes was investigated. Type II cells were isolated either from adult rats with elastase (adult type II cells) or from young rats (4-11 days postnatal) with collagenase and trypsin (young type II cells) and were incubated with dexamethasone (Dex, 10 nM) and cAMP (0.1 mM). By day 4 of culture with hormone treatment, the mRNA levels in adult type II cells were less than 3% of day 0 values, whereas surfactant protein (SP)-A protein content was 26%. However, young type II cells maintained lamellar bodies and microvilli and secreted phospholipid in response to ATP. SP-A, -B, and -C mRNA levels were elevated to 159, 350, and 39%, respectively, of day 0 values with a synergistic response to Dex and cAMP, whereas SP-A protein content rose to 119%. Surfactant mRNA and protein did not recover in cells cultured without hormones. This cell culture system restored surfactant components in rat type II cells.
Bates
Sandra R
SR
The Institute for Environmental Medicine, University of Pennsylvania, 19104, USA. batekenn@mail.med.upenn.edu
Gonzales
Linda W
LW
Tao
Jian-Qin
JQ
Rueckert
Peter
P
Ballard
Philip L
PL
Fisher
Aron B
AB
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D000831:Animals, Newborn; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D017364:Collagenases; D000242:Cyclic AMP / Q000494:pharmacology; D003907:Dexamethasone / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects / Q000502:physiology; D005938:Glucocorticoids / Q000494:pharmacology; D008297:Male; D010196:Pancreatic Elastase; D010641:Phenotype; D010743:Phospholipids / Q000378:metabolism; D011510:Proteolipids / Q000235:genetics* / Q000378:metabolism; D011650:Pulmonary Alveoli / Q000166:cytology* / Q000502:physiology*; D037662:Pulmonary Surfactant-Associated Protein A; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D014357:Trypsin
D005938:Glucocorticoids; D010743:Phospholipids; D011510:Proteolipids; D037662:Pulmonary Surfactant-Associated Protein A; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants; D012333:RNA, Messenger; D003907:Dexamethasone; D000255:Adenosine Triphosphate; D000242:Cyclic AMP; D010196:Pancreatic Elastase; D014357:Trypsin; D017364:Collagenases