带血管肌筋膜包埋脂肪干细胞载体复合物构建血管化组织工程脂肪

doi:10.3969/j.issn.2095-4344.2013.18.017

中国组织工程研究 ›› 2013, Vol. 17 ›› Issue (18): 3349-3357.doi: 10.3969/j.issn.2095-4344.2013.18.017

• 细胞与组织移植 cell and tissue transplantation • 上一篇下一篇

带血管肌筋膜包埋脂肪干细胞载体复合物构建血管化组织工程脂肪

王和庚¹，黎洪棉¹，崔世恩²，徐昆明¹

1中山大学附属中山医院整形美容外科，广东省中山市 528403
2中山大学附属中山医院乳腺外科，广东省中山市 528403

收稿日期:2012-05-31 修回日期:2012-07-24 出版日期:2013-04-30 发布日期:2013-04-30
通讯作者: 黎洪棉，男，1971年生，2008年南方医科大学毕业，博士，主任医师，硕士生导师，主要从事整形美容外科和干细胞、组织工程与再生医学的研究。中山大学附属中山医院整形美容外科，广东省中山市 528403
作者简介:王和庚★，男，1972年生，硕士，副主任医师，主要从事烧伤整形临床与基础研究工作。
基金资助:
课题受中国博士后科学基金(20090450910)，广东省医学科研基金项目(A2011739)和中山市科技计划项目(20113A008)资助。

Vascularized tissue-engineered adipose established via the adipose-derived stem cells-attached scaffolds encapsulated in muscular fasciae with axial pattern blood vessel pedicle

Wang He-geng¹, Li Hong-mian¹, Cui Shi-en², Xu Kun-ming¹

1 Department of Plastic and Aesthetic Surgery, Postdoctoral Mobile Work Station, the Affiliated Zhongshan Hospital of Sun Yat-sen University, Zhongshan 528403, Guangdong Province, China
2 Department of Mammary Gland Surgery, the Affiliated Zhongshan Hospital of Sun Yat-sen University, Zhongshan 528403, Guangdong Province, China

Received:2012-05-31 Revised:2012-07-24 Online:2013-04-30 Published:2013-04-30
Contact: Li Hong-mian, Doctor, Chief physician, Master’s supervisor, Department of Plastic and Aesthetic Surgery, Postdoctoral Mobile Work Station, the Affiliated Zhongshan Hospital of Sun Yat-Sen University, Zhongshan 528403, Guangdong Province, China binrong2112@163.com
About author:Wang He-geng★, Master, Associate chief physician, Department of Plastic and Aesthetic Surgery, the Affiliated Zhongshan Hospital of Sun Yat-Sen University, Zhongshan 528403, Guangdong Province, China drwanghegeng@126.com
Supported by:
China Postdoctoral Science Foundation, No. 20090450910; Guangdong Medical Scientific Foundation, No. A2011739; Zhongshan Scientific and Technological Planning Projects, No. 20113A008

摘要/Abstract

摘要：

背景：再血管化是构建组织工程化脂肪的关键因素。
目的：观察3种不同筋膜瓣包裹的脂肪来源干细胞与胶原蛋白支架复合物在体内成脂效率的差异。
方法：分离兔右侧带血管蒂的背阔肌筋膜瓣，包裹成脂诱导后的脂肪来源干细胞与Ⅰ型胶原蛋白复合物，设为带有轴型血管蒂的诱导分化组。分离兔左侧带血管蒂的背阔肌筋膜瓣，包裹未经诱导的脂肪来源干细胞与Ⅰ型胶原蛋白复合物，设为带有轴型血管蒂的未诱导分化组。分离兔右侧无特定血管蒂的臀大肌筋膜瓣，包裹未经诱导的脂肪来源干细胞与Ⅰ型胶原蛋白复合物，作为对照组。
结果与结论：移植后8周，苏木精-伊红染色和免疫组织化学染色检测结果显示，各组均可见新生脂肪组织形成，带有轴型血管蒂的诱导分化组新生肪组织平均湿质量和新生微血管密度均高于其他2组 (P < 0.01，P < 0.05)。结果说明实验成功构建了带血管肌筋膜包埋成脂诱导后的脂肪干细胞载体复合物构建血管化组织工程脂肪，此复合物在体内的成脂效率和促进血管再生的能力最好。

关键词: 器官移植, 组织移植, 器官移植临床应用, 脂肪干细胞, 成脂分化, 背阔肌筋膜瓣, 臀大肌筋膜瓣, 组织工程化脂肪, 脂肪组织, 脂肪细胞, 血管生成, 细胞移植, 胶原支架, 兔, 组织工程, 省级基金

Abstract:

BACKGROUND: Revascularization mechanism is the decisive factor for the successful construction of tissue-engineered adipose tissue.
OBJECTIVE: To observe the difference of in vivo adipogenic efficacy of adipose-derived stem cells and collagen protein scaffold encapsulated in three muscular fasciae.
METHODS: The rabbit right vascularized latissimus dorsi muscular fasciae was separated and encapsulated with type Ⅰ collagen protein and adipogenic differentiated adipose-derived stem cells complexes as the differentiation group with axial pattern blood vessel pedicle. The rabbit left vascularized latissimus dorsi muscular fasciae was separated and encapsulated with type Ⅰ collagen protein and undifferentiated adipose-derived stem cells complexes as the undifferentiation group with axial pattern blood vessel pedicle. The rabbit gluteus maximus muscular fasciae without specific vascular pedicle was separated and encapsulated with type Ⅰ collagen protein and undifferentiated adipose-derived stem cells complexes as the control group.
RESULTS AND CONCLUSION: Eight weeks after transplantation, hematoxylin-eosin staining and immunohistochemical staining showed that there was new adipose tissue formation in all groups. The mean humid weight of the neonatal fat tissue and the microvessel density in the differentiation group was higher than those in the undifferentiation group and the control group (P < 0.01, P < 0.05). It indicated that the vascularized tissue-engineered adipose was successfully established via adipose-derived stem cells-attached scaffolds encapsulated in muscular fasciae with axial pattern blood vessel pedicle with good in vivo adipogenic efficiency and strong ability to promote angiogenesis.

Key words: organ transplantation, tissue transplantation, clinical application of organ transplantation, adipose-derived stem cells, adipogenic differentiation, latissimus dorsi muscular fasciae, gluteus maximus muscular fasciae, tissue-engineered adipose tissue, adipose tissue, adipose cells, angiogenesis, cell transplantation, collagen scaffold, rabbits, tissue engineering, provincial grants-supported paper

中图分类号:

R318

王和庚，黎洪棉，崔世恩，徐昆明 . 带血管肌筋膜包埋脂肪干细胞载体复合物构建血管化组织工程脂肪[J]. 中国组织工程研究, 2013, 17(18): 3349-3357.

Wang He-geng1, Li Hong-mian1, Cui Shi-en2, Xu Kun-ming1. Vascularized tissue-engineered adipose established via the adipose-derived stem cells-attached scaffolds encapsulated in muscular fasciae with axial pattern blood vessel pedicle[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(18): 3349-3357.

图/表（结果） 4

Quantitative analysis of experimental animals

A total of 10 rabbits involved in the experiment at the beginning. No rabbit dead or got infection until the end of the experiment.

Characterization of adipose-derived stem cells expanded ex vivo

After primary cultured in the control medium, adipose-derived stem cells were harvested with an elongated fibroblast like appearance (Figure 1 a). After expanded to passages 3, the cells were detached and stained with fluorescent dye- 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine (Figure 1 b). The 1,1’-dioctadecyl- 3,3,3',3'- tetramethylindocarbocyanine stained adipose-derived stem cells were selected for the experiment.

General observation and humid weight detection of neogenetic tissue

The histological morphology of the tissues in each group is showed in Figures 2a-c. At 8 weeks after operation, the neogenetic adipose tissue formation was observed macroscopically in the operative area of differentiated group (Figures 2d, g), undifferentiated group (Figures 2e, g) and control group (Figures 2f, g).

And then the hematoxylin-eosin staining showed that the neogenetic tissues were adipose tissues. The mean humid weight was (25.1±1.9) mg of the neogenetic tissue in the differentiated group was higher than that of the undifferentiated group (23.0±1.6) mg and the control group (20.2±1.4) mg (P < 0.01).

Vessel distribution observed with stereomicroscope

Vessels could be seen in the undifferentiated group with axial pattern blood vessel pedicle (Figure 3a) and the differentiated group with axial pattern blood vessel pedicle (Figure 3c). The neogenetic capillaries were found in the undifferentiated group with axial pattern blood vessel pedicle (Figure 3b) and the differentiated group with axial pattern blood vessel pedicle (Figure 3d).

Histological changes of capillary in each group

The scaffolds were degraded thoroughly after 8 weeks at the new formed tissue. Obvious difference could be found about the capillary density in different groups. Compared to the control group (Figure 3f), capillary density of the neogenetic adipose tissue was obviously increased in both the undifferentiated group with axial pattern blood vessel pedicle (Figure 3b) and the adipogenic differentiated group with axial pattern blood vessel pedicle (Figure 3d; P < 0.05). Histological evaluation of 20 fields of neogenetic adipose tissue showed that the mean number of capillaries per field, an index of neovascularization, in the undifferentiated group with axial pattern blood vessel pedicle was (28.5±4.6) capillaries and the adipogenic differentiated group with axial pattern blood vessel pedicle was (31.2±4.5) capillaries, which both had a statistically significant increase in capillary density compared with control group. And control group were (19.3±2.6) capillaries.

Origin of adipocytes in neogenetic adipose tissue and endotheliocytes in neovascularized capillary

The adipocytes and partial vascular endothelial cells presented red fluor in the neogenetic tissue of all groups under fluorescence microscope (Figures 4b, d, f). It indicated that these cells were differentiated from DiI-labeled adipose-derived stem cells. The hematoxylin-eosin stained result proved the structure we observed was capillaries, and the scaffolds were degraded thoroughly (Figures 3b, d, f).

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引言

材料方法

Design

A randomized controlled animal experiment.

Time and setting

The experiment was completed at the Experimental Animal Room and Central Laboratory of the Nanfang Hospital of Southern Medical University from January 2006 to May 2007.

Animals

A total of 10 rabbits aged 3-month-old and weighting 2 500-3 000 g were purchased from Animal Research Institute of China. Rabbits were housed in a light-controlled room with a 12-hour light/dark cycle and had free access to water and food. The experiment was conducted in compliance with the Guidance Suggestions for the Care and Use of Laboratory Animals, formulated by the ministry of Science and Technology of China.

Agents and instruments

Methods

Isolation, expansion and differentiation of rabbit adipose-derived stem cells

Ten rabbits aged three-month-old were used for this study. They were anesthetized with nembutal at 40 mg/kg and shaved. The inguinal fat pads were harvested by exairesis from the rabbits (The Southern Medical University institutional ethics committee approved all experiments in advance).

The tissue was washed at least three times with Phosphate buffered saline (Sigma) to remove contaminating blood, and then excised, finely minced. Adipose tissue samples were digested at 37 ℃ for 60 minutes with intermittent shaking in 0.1% (w/v) collagenase typeⅠ(Gibco-BRL, Gaithersburg, MD, USA). The floating adipocytes were separated from the stromal-vascular fraction by centrifugation at 260 g for 5 minutes. The isolated stem cells in the stromal-vascular fraction were cultured in Dulbecco’s modified Eagle’s medium-high glucose medium supplemented with 10% (v/v) fetal bovine serum and maintained for 1-3 days. After this period, the culture flasks were washed with phosphate-buffered saline to remove any non-adherent cells and fed with an expansion medium consisting of Dullbecco’s modified Eagle’s medium-high glucose medium supplemented with 10% (v/v) fetal bovine serum. Cells were expanded for 7-9 days until they achieved confluence. Cultures were observed with a microscope to assess expansion and cell morphology.

In order to prevent differentiation, cells were harvested at 80% to 90% confluent and then the cultured cells were detached from the culture dishes with 0.25% trypsin, neutralized with culture medium, and collected by centrifugation at 200 g for 5 minutes at room temperature. Cells were then passaged 1:3 per 3 to 4 days. Only cultured cells for 3 passages were analyzed in this study. The cells were suspended in medium at the concentration of 1×10⁷ cells per 1 mL for co-culture with collagen type Ⅰ scaffolds. On the other hand, confluent cultured stem cells were induced to undergo adipogenesis with the following differentiation medium: Dulbecco’s modified Eagle’s medium-high glucose supplemented with 10% (v/v) fetal bovine serum, human recombinant, insulin (10 μmol/L), dexamethasone (1 μmol/L), isobutylmethylxanthine (0.5 mmol/L) and indomethacin (0.2 mmol/L). After a 3-day induction period, the cells were fed with the same medium, every 2-3 days for the remaining culture period. The adipogenic differentiation of the stem cells was confirmed by light microscopic observation and the morphological changes from fibroblast-like structures to round cell structures with lipid vacuoles were observed with oil red O staining.

Oil red O staining

Following 10-12 days of differentiation into adipocytes, cells were fixed in 10% (v/v) neutral buffered formaldehyde for 1 hour and stained for 10 minutes with a filtered 60% (v/v) oil red O (Sigma) solution in distilled water. Then, cells were washed with distilled water to remove remaining oil red O solution. To determine the rate of adipogenic differentiation, detached cells were also stained with oil red O. After cells detached using trypsin/ethylenediaminetetraacetic acid solution [0.1% (w/v) trypsin and 0.08 g/L ethylene diamine tetraacetic acid], the resuspended cells were fixed in 10% (v/v) neutral buffered formaldehyde for 1 hour and stained for 10 minutes with 60% (v/v) oil red O solution. The differentiation rate was expressed as the ratio of the number of oil red O-positive cells to the number of total cells. Undifferentiated stem cells were also stained with oil red O.

Adipose-derived stem cells labeled with dissolve 1,1'-dioctadecyl-3,3,3’,3’- tetramethylindocarbocyanine

1,1'-dioctadecyl-3,3,3',3'- tetramethylindocarbocyanine (Molecular Probes, Eugene, Ore.) was treated with 99% ethanol in the concentration of 25% then stored at -20 ℃ for application. Adipose-derived stem cells were labeled with fluorescent 1,1'-dioctadecyl- 3,3,3',3'- tetramethylindocarbocyanine according to the manufacturer’s recommendations; and cells in suspension were incubated with 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine at the concentration of 2.5 mg/L in phosphate-buffered saline for 5 minutes at 37 ℃ and for 15 minutes at 4 ℃.

Establishment of latissimus dorsi myocutaneous flap model

The double side portait latissimus dorsi myocutaneous flaps were produced on the back which located the two side of spinal median line. The length breadth ratio is

20 mm:50 mm, their axial vessels were thoracodorsal artery and thoracodorsal vein. A cycloid with 1.5 cm diameter sac was separated in fascia musculares of the latissimus dorsi myocutaneous flap.

Grouping and intervention

The experiment was divided into three groups. The undifferentiated group with axial pattern blood vessel pedicle (n=10). The differentiated group with axial pattern blood vessel pedicle (n=10). Control group (n=10). In the undifferentiated group with axial pattern blood vessel pedicle, one piece of type Ⅰ collagen sponge loaded with normal adipose-derived stem cells (1×10⁷ cell population) was transplanted into the fascia musculares sac with blood vessel pedicle of left latissimus dorsi myocutaneous flaps; and in the differentiated group with axial pattern blood vessel pedicle, the same size collagen sponge loaded with the adipogenic differentiated adipose-derived stem cells were transplanted into the fascia musculares sac with axial pattern blood vessel pedicle of right latissimus dorsi myocutaneous flaps at the same rabbits. In the differentiated group without axial pattern blood vessel pedicle (control group), another same size collagen sponge loaded with adipogenic differentiated adipose-derived stem cells were also implanted into the random fascia musculares sac without blood vessel pedicle of right gluteus myocutaneous flaps. The rabbits were fed routinely after operation.

General observation and mean humid weight detection of neogenetic tissue

Euthanasia was performed at the animals at 8 weeks after transplantation of adipose-derived stem cells with scaffolds. No animal from either the study or control group died during the study period. The neogenetic tissue were observed macroscopically, then they were dissected out and weighed by electronic balance (fidelity=0.000 1 g).

Vessel distribution observed with stereomicroscope and the neovascularized capillary density observed with hematoxylin-eosin staining

Tissue sections from neogenetic tissue of each group were harvested on the postoperative 8 weeks. Vessels distributions were observed with stereomicroscope and the tissue were embedded in paraffin for conventional hematoxylin-eosin staining histological assessment.

Neovascularization was assessed by measuring the number of capillaries in 10 fields of hematoxylin-eosin staining slide (100 magnification; measurements were performed by two blinded reviewers).

Origin of adipocytes in neogenetic adipose tissue and endotheliocytes in neovascularized capillary

We labeled cell membrane of the adipose-derived stem cells with 1,1'-dioctadecyl-3,3,3',3'- tetramethylindocarbocyanine before transplant the adipose-derived stem cells-attached typeⅠcollagen scaffolds complex into the animals’ body. The grafts were dislodged after 8 weeks. The cells contained 1,1'-dioctadecyl- 3,3,3',3'-tetramethylindocarbocyanine if the neogenetic adipocytes and vascular endothelial cells differentiated from the adipose-derived stem cells which we have implanted. Therefore, the adipocytes and the vascular endothelial cells presented red fluor in the neogenetic tissue, so it can be thought that the red adipocytes and vascular endotheliocytes were differentiated from exogenous adipose-derived stem cells.

Main outcome measures

The humid weight of the neogenetic tissues in all groups, distribution and density of micro newborn adipose microvascular, morphology of blood capillary and morphology of vascularized tissue-engineered adipose were observed.

Statistical analysis

All data were analyzed using SPSS for Windows 10.0 software (SPSS, Chicago, IL, USA) and presented as mean±SD. One-way analysis of variance was used to compare the means for three groups simultaneously. Furthermore, Post Hoc tests were performed to examine the difference between any two groups, using least significant difference or Tamhane method when variances were not homogenous across groups. The difference was regarded statistically significant if a two-tail P value was less than 0.05.

讨论

There are two types of “true” totipotent stem cells, both of which are derived from embryonic sources. One is embryonic stem cells, and the other type of totipotent stem cells is embryonic germ cells. Embryonic stem/embryonic germ cells have the potential to differentiate into all kinds of cells. However, there are problems associated with the clinical use of embryonic stem/embryonic germ cells^[17]. The first and most obvious problem is ethical concerns. The second is immunologic rejection by recipients. The third problem is the risk of tumor formation, especially teratomas or teratocarcinomas, resulting from contamination of uncontrolled cells.

Compared to embryonic stem/embryonic germ cells, applications of adult stem cells face few ethical problems. In addition, immunologic problems do not have to be considered in auto transplantation. Among them, bone marrow-derived stem cells are the most notable type with kinds of multipotency since they were first described in 1976^[18]. Bone marrow-derived stem cells were testified to have the potency to differentiate not only into paraxial and visceral mesoderm cells, but also into neuroectoderm and endoderm cells^[19-21].

Bone marrow is the major source of bone marrow-derived stem cells for a long time, but recent studies have identified new sources. Zuk et al ^[22]have found bone marrow-derived stem cells in adipose tissue. At the 2^ndAnnual Meeting of the International Fat Applied Technology Society (IFATS, Pittsburgh, 2004), nomenclature was discussed and it was concluded that the term “adipose derived stem cells” or “ASCs” should be used for pluripotent cells harvested from adipose tissue. Adipose-derived stem cells are ideal in many aspects: they can be harvested, handled and multiplied non-invasively, easily, and effectively; their pluripotency and proliferative efficiency are not less than those of bone marrow-derived bone marrow-derived stem cells; and donor morbidity is lower than that of bone marrow-derived stem cells harvested from other sites.

Engineered compound tissue is regarded as the combination of cells and scaffold constructed in vitro. For the effective construction of engineered-tissue in vivo, we must ensure the seed cells, especially the cells in the scaffolds, to be nourished promptly and sufficiently. Therefore, the re-establishment of blood supply is the key to tissue engineering^[23-24].

Improving the viability of seed cells depends on increased neovascularization which occurs through two distinct mechanisms of action. The first mechanism is angiogenesis and the other is called vasculogenesis. Through the general observation, we have obviously detected the increased capillary density in the undifferentiated group with axial pattern blood vessel pedicle and the adipogenic differentiated group with axial pattern blood vessel pedicle, which were also proved by following histological assessments. These increased capillaries may directly account for the increasing of adipose-derived stem cells’ viability and humid weight of the neogenetic adipose tissue in undifferentiated group and differentiated group.

In order to clarify the mechanism of the angiogenesis or vasculogenesis, we tried to detect the origin of these increased capillary. In another word, are these increased capillary vessels originated from preexisting “blocked” capillary or they were newly formed? In this experiment, we used fluorescent positive 1,1'-dioctadecyl-3,3,3',3'- tetramethylindocarbocyanine staining to label the adipose-derived stem cells. With the help of fluorescent observation, we revealed that partial endothelial in experimental groups were positive (both undifferentiated group and differentiated group). Endothelial cells positively for 1,1'-dioctadecyl-3,3,3',3'- tetramethylindocarbocyanine indicated that these cells were differentiated from administrated adipose-derived stem cells. These results indicated that at least parts of the endothelial were from exogenously transplanted adipose-derived stem cells and newly differentiated endothelial may participated in the construction of capillary. The rest endothelial cells negatively for 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine indicated that these cells were preexisting “blocked” ones which originated from the axial pattern vessels pedicle of rabbits’ latissimus dorsi muscle. These results indicated that increased neovascularization which occurs both through two distinct mechanisms of action in the experimental groups. However, the increased neovascularization which occurs only through one mechanisms of action in the control group, because its adipose-derived stem cells-attached type Ⅰ collagen scaffolds were transplanted into the fascial flap of gluteus maximus without axial pattern blood vessel pedicle. So it can account for the improvement of increased the adipose-derived stem cells’ viability and humid weight of the neogenetic adipose tissue in the undifferentiated group with axial pattern blood vessel pedicle and the adipogenic differentiated group with axial pattern blood vessel pedicle.

Skin flap, musculocutaneous flap, muscle flap and fascial flap with vessel pedicle are pervasively utilized to repair the defect of various soft tissues or organs in clinic. Accordingly, we can carry out the vascularization of engineered adipose tissue by encapsulating in musculocutaneous flap, muscle flap or fascial flap with blood supply^[25-27]. Muscular fasciae with vessel pedicle are nourished with abundant blood supply. On account of the property, we encapsulated the adipose-derived stem cells-attached typeⅠcollagen scaffolds complex of adipogenic undifferentiation-inducing or adipogenic differentiation-inducing conditions by latissimus dorsi fascia with its vessel pedicle. After 8 weeks, histological detection showed that the neogenetic tissue is adipose tissue in all groups. Though, we also found a certain neogenetic adipose tissue in the control group. However, both the mean humid weight and neogenetic capillary vessel density in the undifferentiated group with axial pattern blood vessel pedicle group and the adipogenic differentiated group with axial pattern blood vessel pedicle group are significantly larger than those in the control group. These demonstrate that muscular fasciae flap with vessel pedicle can preferably facilitate the survival of seed cells and promote the neovascularization of the grafts. So it can fabricate a larger volume engineered adipose tissue than that of the adipose-derived stem cells-attached typeⅠcollagen scaffolds complex of adipogenic differentiation-inducing encapsulated by muscular fasciae without vessel pedicle.

In the past, some scholars thought that the undifferentiated inducing stem cells which transplanted in vivo could not differentiate into reciprocal terminal cells. But in our study, after 8 weeks of the transplantation, the novel phenomenon is that neogenetic adipose tissues formation could be observed when the adipogenic undifferentiated adipose-derived stem cells-attached with scaffolds transplanted into animals’ body. The possible mechanism is that the adipose-derived stem cells which we implanted are a stem cell colonia, which may contains some preadipocytes. These preadipocytes would be differentiated into adipocytes when they were transplanted into human body.

In this study, conclusion can be drawn that the adipose-derived stem cells-attached scaffolds encapsulated in muscular fasciae with axial pattern blood vessel pedicle can increase the neovascularization and enhance the tissue-engineering adipogenesis. And future’s studies will be underwent to help to determine the optimal mode and timing of adipose-derived stem cells therapy for therapeutic efficacy, and to investigate the potential of these cells to act as tissue-engineering adipogenesis for the reconstruction or augmentation of soft tissues deficiency due to mastectomy or micromastia in plastic and reconstructive or aesthetic surgery.

This study demonstrates that the transplantation of autologous adipose-derived stem cells-attached scaffolds encapsulated in muscular fasciae with axial pattern blood vessel pedicle can increase the survival rate of adipose-derived stem cells and enhance the tissue-engineering adipogenesis. Autologous adipose-derived stem cells can be differentiated into endothelial cells and then increase the neovascularization.

带血管肌筋膜包埋脂肪干细胞载体复合物构建血管化组织工程脂肪

Vascularized tissue-engineered adipose established via the adipose-derived stem cells-attached scaffolds encapsulated in muscular fasciae with axial pattern blood vessel pedicle

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