Human breast adipose‑derived stem cells: Characterization and differentiation into mammary gland‑like epithelial cells promoted by autologous activated platelet‑rich plasma

Cui,Shi-En; Li,Hong-Mian; Liu,Da-Lie; Nan,Hua; Xu,Kun-Ming; Zhao,Pei-Ran; Liang,Shuang-Wu

doi:10.3892/mmr.2014.2280

Human breast adipose‑derived stem cells: Characterization and differentiation into mammary gland‑like epithelial cells promoted by autologous activated platelet‑rich plasma

Authors:
- Shi-En Cui
- Hong-Mian Li
- Da-Lie Liu
- Hua Nan
- Kun-Ming Xu
- Pei-Ran Zhao
- Shuang-Wu Liang
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Affiliations: Department of Mammary Gland Surgery, Zhongshan Hospital of Sun Yat‑Sen University, Zhongshan, Guangdong 528403, P.R. China, Department of Plastic and Aesthetic Surgery, Zhongshan Bo'ai Hospital of Southern Medical University, Zhongshan, Guangdong 528403, P.R. China, Department of Plastic and Reconstructive Surgery, Zhujiang Hospital of Southern Medical University, Guangzhou, Guangdong 510282, P.R. China, Research Center for Tissue Engineering, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China
Published online on: May 28, 2014 https://doi.org/10.3892/mmr.2014.2280
Pages: 605-614

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Abstract

Human adipose‑derived stem cells (ASCs) isolated from various body sites have been widely investigated in basic and clinical studies. However, ASCs derived from human breast tissue (hbASCs) have not been extensively investigated. In order to expand our understanding of hbASCs and examine their potential applications in stem cell research and cell‑based therapy, hbASCs were isolated from discarded surgical fat tissue following reduction mammoplasty and a comprehensive characterization of these hbASCs was performed, including analysis of their cellular morphology, growth features, cell surface protein markers and multilineage differentiation capacity. These hbASCs expressed cluster of differentiation (CD)44, CD49d, CD90 and CD105, but did not express CD31 and CD34. Subsequently, the hbASCs were differentiated into adipocytes, osteocytes and chondrocytes in vitro. In order to examine the potential applications of hbASCs in breast reconstruction, an approach to promote in vitro differentiation of hbASCs into mammary gland‑like epithelial cells (MGECs) was developed using activated autologous platelet‑rich plasma (PRP). A proliferation phase and a subsequent morphological conversion phase were observed during this differentiation process. PRP significantly promoted the growth of hbASCs in the proliferation phase and increased the eventual conversion rate of hbASCs into MGECs. Thus, to the best of our knowledge, the present study provided the first comprehensive characterization of hbASCs and validated their multipotency. Furthermore, it was revealed that activated autologous PRP was able to enhance the differentiation efficiency of hbASCs into MGECs. The present study and other studies of hbASCs may aid the development of improved breast reconstruction strategies.

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1	Cordeiro PG: Breast reconstruction after surgery for breast cancer. N Engl J Med. 359:1590–1601. 2008. View Article : Google Scholar : PubMed/NCBI
2	Ren G, Chen X, Dong F, et al: Concise review: mesenchymal stem cells and translational medicine: emerging issues. Stem Cells Transl Med. 1:51–58. 2012. View Article : Google Scholar : PubMed/NCBI
3	Dubois SG, Floyd EZ, Zvonic S, Kilroy G, Wu X, Carling S, Halvorsen YD, Ravussin E and Gimble JM: Isolation of human adipose-derived stem cells from biopsies and liposuction specimens. Methods Mol Biol. 449:69–79. 2008.PubMed/NCBI
4	Neupane M, Chang CC, Kiupel M and Yuzbasiyan-Gurkan V: Isolation and characterization of canine adipose-derived mesenchymal stem cells. Tissue Eng Part A. 14:1007–1015. 2008. View Article : Google Scholar : PubMed/NCBI
5	Peptan IA, Hong L and Mao JJ: Comparison of osteogenic potentials of visceral and subcutaneous adipose-derived cells of rabbits. Plast Reconstr Surg. 117:1462–1470. 2006. View Article : Google Scholar : PubMed/NCBI
6	Tholpady SS, Katz AJ and Ogle RC: Mesenchymal stem cells from rat visceral fat exhibit multipotential differentiation in vitro. Anat Rec A Discov Mol Cell Evol Biol. 272:398–402. 2003. View Article : Google Scholar : PubMed/NCBI
7	Vidal MA, Kilroy GE, Lopez MJ, Johnson JR, Moore RM and Gimble JM: Characterization of equine adipose tissue-derived stromal cells: adipogenic and osteogenic capacity and comparison with bone marrow-derived mesenchymal stromal cells. Vet Surg. 36:613–622. 2007. View Article : Google Scholar
8	Safford KM, Hicok KC, Safford SD, et al: Neurogenic differentiation of murine and human adipose-derived stromal cells. Biochem Biophys Res Commun. 294:371–379. 2002. View Article : Google Scholar : PubMed/NCBI
9	Jurgens WJ, Oedayrajsingh-Varma MJ, Helder MN, Zandiehdoulabi B, Schouten TE, Kuik DJ, Ritt MJ and van Milligen FJ: Effect of tissue-harvesting site on yield of stem cells derived from adipose tissue: implications for cell-based therapies. Cell Tissue Res. 332:415–426. 2008. View Article : Google Scholar : PubMed/NCBI
10	Grimes BR, Steiner CM, Merfeld-Clauss S, et al: Interphase FISH demonstrates that human adipose stromal cells maintain a high level of genomic stability in long-term culture. Stem Cells Dev. 18:717–724. 2009. View Article : Google Scholar : PubMed/NCBI
11	Zuk PA, Zhu M, Ashjian P, et al: Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell. 13:4279–4295. 2002.PubMed/NCBI
12	Gimble JM and Guilak F: Differentiation potential of adipose derived adult stem (ADAS) cells. Curr Top Dev Biol. 58:137–160. 2003. View Article : Google Scholar : PubMed/NCBI
13	Gimble J and Guilak F: Adipose-derived adult stem cells: isolation, characterization, and differentiation potential. Cytotherapy. 5:362–369. 2003. View Article : Google Scholar : PubMed/NCBI
14	Strem BM, Hicok KC, Zhu M, et al: Multipotential differentiation of adipose tissue-derived stem cells. Keio J Med. 54:132–141. 2005. View Article : Google Scholar : PubMed/NCBI
15	Fraser JK, Schreiber R, Strem B, et al: Plasticity of human adipose stem cells toward endothelial cells and cardiomyocytes. Nat Clin Pract Cardiovasc Med. 3(Suppl 1): S33–S37. 2006. View Article : Google Scholar : PubMed/NCBI
16	Jack GS, Almeida FG, Zhang R, Alfonso ZC, Zuk PA and Rodríguez LV: Processed lipoaspirate cells for tissue engineering of the lower urinary tract: implications for the treatment of stress urinary incontinence and bladder reconstruction. J Urol. 174:2041–2045. 2005. View Article : Google Scholar
17	Rodríguez LV, Alfonso Z, Zhang R, Leung J, Wu B and Ignarro LJ: Clonogenic multipotent stem cells in human adipose tissue differentiate into functional smooth muscle cells. Proc Natl Acad Sci USA. 103:12167–12172. 2006.PubMed/NCBI
18	Mizuno H, Zuk PA, Zhu M, Lorenz HP, Benhaim P and Hedrick MH: Myogenic differentiation by human processed lipoaspirate cells. Plast Reconstr Surg. 109:199–209. 2002. View Article : Google Scholar : PubMed/NCBI
19	Zaman WS, Makpol S, Sathapan S and Chua KH: Long-term in vitro expansion of human adipose-derived stem cells showed low risk of tumourigenicity. J Tissue Eng Regen Med. 8:67–76. 2014. View Article : Google Scholar : PubMed/NCBI
20	Lee JH and Kemp DM: Human adipose-derived stem cells display myogenic potential and perturbed function in hypoxic conditions. Biochem Biophys Res Commun. 341:882–888. 2006. View Article : Google Scholar : PubMed/NCBI
21	Liu TM, Martina M, Hutmacher DW, Hui JH, Lee EH and Lim B: Identification of common pathways mediating differentiation of bone marrow- and adipose tissue-derived human mesenchymal stem cells into three mesenchymal lineages. Stem Cells. 25:750–760. 2007.
22	Li HX, Luo X, Liu RX, Yang YJ and Yang GS: Roles of Wnt/beta-catenin signaling in adipogenic differentiation potential of adipose-derived mesenchymal stem cells. Mol Cell Endocrinol. 291:116–124. 2008. View Article : Google Scholar : PubMed/NCBI
23	Karbiener M, Fischer C, Nowitsch S, et al: microRNA miR-27b impairs human adipocyte differentiation and targets PPARgamma. Biochem Biophys Res Commun. 390:247–251. 2009. View Article : Google Scholar : PubMed/NCBI
24	James AW, Leucht P, Levi B, et al: Sonic Hedgehog influences the balance of osteogenesis and adipogenesis in mouse adipose-derived stromal cells. Tissue Eng Part A. 16:2605–2616. 2010. View Article : Google Scholar : PubMed/NCBI
25	Davis LA and Zur Nieden NI: Mesodermal fate decisions of a stem cell: the Wnt switch. Cell Mol Life Sci. 65:2658–2674. 2008. View Article : Google Scholar : PubMed/NCBI
26	Schäffler A and Buchler C: Concise review: adipose tissue-derived stromal cells - basic and clinical implications for novel cell-based therapies. Stem Cells. 25:818–827. 2007.PubMed/NCBI
27	Cousin B, André MM, Arnaud E, Pénicaud L and Casteilla L: Reconstitution of lethally irradiated mice by cells isolated from adipose tissue. Biochem Biophys Res Commun. 301:1016–1022. 2003. View Article : Google Scholar : PubMed/NCBI
28	Miranville A, Heeschen C, Sengenès C, Curat CA, Busse R and Bouloumié A: Improvement of postnatal neovascularization by human adipose tissue-derived stem cells. Circulation. 110:349–355. 2004. View Article : Google Scholar : PubMed/NCBI
29	Corre J, Barreau C, Cousin B, et al: Human subcutaneous adipose cells support complete differentiation but not self-renewal of hematopoietic progenitors. J Cell Physiol. 208:282–288. 2006. View Article : Google Scholar : PubMed/NCBI
30	Li H, Han Z, Liu D, Zhao P, Liang S and Xu K: Autologous platelet-rich plasma promotes neurogenic differentiation of human adipose-derived stem cells in vitro. Int J Neurosci. 123:184–190. 2013. View Article : Google Scholar : PubMed/NCBI
31	Wu J, Pan Z, Cheng M, et al: Ginsenoside Rg1 facilitates neural differentiation of mouse embryonic stem cells via GR-dependent signaling pathway. Neurochem Int. 62:92–102. 2013. View Article : Google Scholar : PubMed/NCBI
32	Shi AW, Gu N, Liu XM, Wang X and Peng YZ: Ginsenoside Rg1 enhances endothelial progenitor cell angiogenic potency and prevents senescence in vitro. J Int Med Res. 39:1306–1318. 2011. View Article : Google Scholar : PubMed/NCBI
33	Wang L and Kisaalita WS: Administration of BDNF/ginsenosides combination enhanced synaptic development in human neural stem cells. J Neurosci Methods. 194:274–282. 2011. View Article : Google Scholar : PubMed/NCBI
34	Shi AW, Wang XB, Lu FX, Zhu MM, Kong XQ and Cao KJ: Ginsenoside Rg1 promotes endothelial progenitor cell migration and proliferation. Acta Pharmacol Sin. 30:299–306. 2009. View Article : Google Scholar : PubMed/NCBI
35	Moshtagh PR, Emami SH and Sharifi AM: Differentiation of human adipose-derived mesenchymal stem cell into insulin-producing cells: an in vitro study. J Physiol Biochem. 69:451–458. 2013. View Article : Google Scholar : PubMed/NCBI
36	Scuderi N, Ceccarelli S, Onesti MG, et al: Human adipose-derived stromal cells for cell-based therapies in the treatment of systemic sclerosis. Cell Transplant. 22:779–795. 2013. View Article : Google Scholar : PubMed/NCBI
37	Kisiel AH, McDuffee LA, Masaoud E, Bailey TR, Esparza Gonzalez BP and Nino-Fong R: Isolation, characterization, and in vitro proliferation of canine mesenchymal stem cells derived from bone marrow, adipose tissue, muscle, and periosteum. Am J Vet Res. 73:1305–1317. 2012. View Article : Google Scholar : PubMed/NCBI
38	Heneidi S, Simerman AA, Keller E, Singh P, Li X, Dumesic DA and Chazenbalk G: Awakened by cellular stress: isolation and characterization of a novel population of pluripotent stem cells derived from human adipose tissue. PLoS One. 8:e647522013. View Article : Google Scholar : PubMed/NCBI
39	Hanson SE, Kim J and Hematti P: Comparative analysis of adipose-derived mesenchymal stem cells isolated from abdominal and breast tissue. Aesthet Surg J. 33:888–898. 2013. View Article : Google Scholar : PubMed/NCBI
40	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods. 25:402–408. 2001.
41	Smith SE and Roukis TS: Bone and wound healing augmentation with platelet-rich plasma. Clin Podiatr Med Surg. 26:559–588. 2009. View Article : Google Scholar : PubMed/NCBI
42	Gentile P, Orlandi A, Scioli MG, Di Pasquali C, Bocchini I and Cervelli V: Concise review: adipose-derived stromal vascular fraction cells and platelet-rich plasma: basic and clinical implications for tissue engineering therapies in regenerative surgery. Stem Cells Transl Med. 1:230–236. 2012. View Article : Google Scholar
43	Mardani M, Kabiri A, Esfandiari E, Esmaeili A, Pourazar A, Ansar M and Hashemibeni B: The effect of platelet rich plasma on chondrogenic differentiation of human adipose derived stem cells in transwell culture. Iran J Basic Med Sci. 16:1163–1169. 2013.PubMed/NCBI
44	Gaiba S, França LP, França JP and Ferreira LM: Characterization of human adipose-derived stem cells. Acta Cir Bras. 27:471–476. 2012. View Article : Google Scholar : PubMed/NCBI
45	Mafi P, Hindocha S, Mafi R, Griffin M and Khan WS: Adult mesenchymal stem cells and cell surface characterization - a systematic review of the literature. Open Orthop J. 5:253–260. 2011. View Article : Google Scholar : PubMed/NCBI
46	Hass R, Kasper C, Böhm S and Jacobs R: Different populations and sources of human mesenchymal stem cells (MSC): A comparison of adult and neonatal tissue-derived MSC. Cell Commun Signal. 9:122011.PubMed/NCBI
47	Dimri G, Band H and Band V: Mammary epithelial cell transformation: insights from cell culture and mouse models. Breast Cancer Res. 7:171–179. 2005. View Article : Google Scholar : PubMed/NCBI
48	Péchoux C, Gudjonsson T, Ronnov-Jessen L, Bissell MJ and Petersen OW: Human mammary luminal epithelial cells contain progenitors to myoepithelial cells. Dev Biol. 206:88–99. 1999.PubMed/NCBI
49	Tobita M, Orbay H and Mizuno H: Adipose-derived stem cells: current findings and future perspectives. Discov Med. 11:160–170. 2011.PubMed/NCBI
50	Zuk PA, Zhu M, Mizuno H, et al: Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng. 7:211–228. 2001. View Article : Google Scholar : PubMed/NCBI
51	Halvorsen YD, Franklin D, Bond AL, et al: Extracellular matrix mineralization and osteoblast gene expression by human adipose tissue-derived stromal cells. Tissue Eng. 7:729–741. 2001. View Article : Google Scholar : PubMed/NCBI
52	Baer PC, Döring C, Hansmann ML, Schubert R and Geiger H: New insights into epithelial differentiation of human adipose-derived stem cells. J Tissue Eng Regen Med. 7:271–278. 2011. View Article : Google Scholar : PubMed/NCBI
53	Baer PC, Brzoska M and Geiger H: Epithelial differentiation of human adipose-derived stem cells. Methods Mol Biol. 702:289–298. 2011. View Article : Google Scholar : PubMed/NCBI