[In vitro study on proliferation and multi-lineage differentiation potential of adipose-derived cells].
Adipose were obtained from patients underwent liposuction treatment (total 19 female donors, 31.5 +/- 5.8 years old). Liposuction tissues were digested with type I collagenase, cells were isolated and cultured up to passage 10. To evaluate the proliferation potential of ADCs, growth curve and cumulative population doubling were achieved by cell counting. CD29,CD105, CD106, CD166, CD49d, CD34, CD31, 3G5 were analyzed by flow cytometry and immunocytochemistry to characterize the cell population. The multi-lineage potential of ADCs was testified by differentiating cells with osteogenic,chondrogenic and adipogenic inducer. A total of 5x10(7) nucleared cells could be obtained from 300ml liposuction tissues. After in vitro cultivation,cumulative population doubling number reached 15.53 at passage 10 (average 1.59 +/- 0.224 /passage). Flow cytometry and immunocytochemistry showed that ADCs expressed high level (>60%) of stem cell-related antigen (CD29, CD105, CD106, CD166), while cells expressed hematopoiesis-related antigen CD34 and CD31 around 7.3% and 29.2% respectively. Collagen II (both in mRNA and protein level) was detected in chondrogenic differentiation. The calcified nodules were observed by von Kossa and Alizarn Red staining and the expressions of AKP and Osteonectin were detected by RT-PCR in osteogenic differentiation. PPARr2, GLU-4, and Leptin genes were detected in adipogenic differentiation and intracellular lipid droplets could be observed by Oil Red staining. ADCs can be abundantly harvested and have high proliferative and multi-lineage differentiation potential.
Zhang
Ying
Y
Department of Plastic and Reconstructive Surgery, Shanghai 9th People's Hospital, Shanghai Second Medical University.
Zhou
Guang Dong
GD
Yang
Ping
P
Yin
Shuo
S
Liu
De Li
DL
Cui
Lei
L
Liu
Wei
W
Cao
Yi Lin
YL
D017667:Adipocytes / Q000166:cytology* / Q000378:metabolism; D000328:Adult; D015703:Antigens, CD / Q000032:analysis; D018952:Antigens, CD34 / Q000032:analysis; D015816:Cell Adhesion Molecules, Neuronal / Q000032:analysis; D002454:Cell Differentiation / Q000502:physiology*; D019070:Cell Lineage; D049109:Cell Proliferation; D002478:Cells, Cultured; D000071063:Endoglin; D005260:Female; D005326:Fetal Proteins / Q000032:analysis; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D039441:Integrin alpha4 / Q000032:analysis; D019012:Integrin beta1 / Q000032:analysis; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000032:analysis; D011956:Receptors, Cell Surface / Q000032:analysis; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D019010:Vascular Cell Adhesion Molecule-1 / Q000032:analysis; D020410:Activated-Leukocyte Cell Adhesion Molecule
D015703:Antigens, CD; D018952:Antigens, CD34; D015816:Cell Adhesion Molecules, Neuronal; D000071063:Endoglin; D005326:Fetal Proteins; D039441:Integrin alpha4; D019012:Integrin beta1; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D011956:Receptors, Cell Surface; D019010:Vascular Cell Adhesion Molecule-1; C105613:ALCAM protein, human; D020410:Activated-Leukocyte Cell Adhesion Molecule; C064206:ENG protein, human