脂肪组织来源干细胞在皮肤与脂肪组织再生中作用的研究
发布时间：2018-08-01 12:59
【摘要】：外伤、感染、糖尿病溃疡以及肿瘤切除术后等常常造成皮肤和脂肪等软组织的缺损,软组织缺损的修复问题长期困扰着临床医生。目前临床上的治疗手段存在各种各样的缺点,随着组织工程和干细胞技术的发展与应用,利用干细胞结合生物材料构建皮肤、脂肪等工程化软组织成为修复软组织缺损最有前景的方法。脂肪组织来源干细胞是一类来源于脂肪组织的成体间充质干细胞,具有自我更新和多向分化的能力,并且具有取材容易、取材量大、损伤较小、细胞增殖快速、干细胞含量丰富等优点,是组织工程理想的种子细胞。本课题首先利用分离的SD大鼠脂肪组织来源干细胞,在鼠的皮肤缺损模型中证明其具有促进创面愈合的能力,并在此基础上进一步分离获取人的脂肪组织来源干细胞,并与真皮成纤维细胞混合模拟真皮微环境,探讨人脂肪组织来源干细胞在组织工程皮肤制备中的作用;另外将人脂肪组织来源干细胞结合可注射材料明胶微球,制备工程化脂肪组织。通过基于脂肪组织来源干细胞构建的组织工程皮肤和脂肪,探索修复软组织缺损的新策略。 本课题主要进行了以下几方面的研究: 1大鼠脂肪组织来源干细胞在皮肤创面愈合中的应用 目的:体外分离培养SD大鼠脂肪组织来源干细胞,对其进行生物学特性研究,并接种于胶原凝胶中构建组织工程真皮修复大鼠皮肤缺损,探讨其作为组织工程皮肤种子细胞的可行性。方法:分离SD大鼠腹股沟部脂肪组织,采用胶原酶消化获得脂肪组织来源干细胞,用MTT法检测细胞的生长特性,流式细胞仪检测细胞周期和细胞表面分子标志物,通过诱导分化检测脂肪组织来源干细胞的多向分化潜能,在验证获取的脂肪组织来源干细胞是间充质干细胞的基础上,进行Hoechst33342荧光标记并与胶原凝胶混合制备组织工程真皮,用于修复大鼠皮肤缺损,随机分为3组:脂肪组织来源干细胞复合胶原移植组(ADSC+Col),单纯胶原移植组(Col),空白对照组(Cont.),并通过组织学和愈合速度检测修复效果。结果:分离培养的脂肪组织来源干细胞具有不断增殖的能力,表达间充质干细胞的表面标志物,具有向脂肪、骨和神经等多个谱系分化的能力;在复合胶原凝胶修复皮肤缺损实验中,脂肪组织来源干细胞复合胶原移植组(ADSC+Col)、单纯胶原移植组(Col)、空白对照组(Cont.)创面愈合时间分别为:(14.3±1.7),(16.9±2.5)和(21.2±4.2)天,有显著性差异(P0.05)。组织学观察显示,脂肪组织来源干细胞复合胶原移植组,光镜下可见大量成纤维细胞和真皮胶原基质长入创面,同时上皮覆盖创面,已形成钉突样结构,但是未见毛囊和汗腺等皮肤附属器结构;单纯胶原移植组次之,上皮爬行但未完全覆盖创面;空白对照组真皮层中微血管较多,但是上皮爬行速度慢,未覆盖创面。标记Hoechst33342的脂肪组织来源干细胞复合胶原移植2周后,可见蓝色荧光的细胞核,证明移植的脂肪组织来源干细胞存活于创面。结论:成功分离了大鼠脂肪组织来源干细胞,脂肪组织来源干细胞复合胶原构建的组织工程真皮,可以修复皮肤软组织缺损,促进创面愈合,证明脂肪组织来源干细胞作为组织工程皮肤构建的种子细胞的可行性。 2人脂肪组织来源干细胞在组织工程皮肤制备中的作用 目的:体外分离培养人脂肪组织来源干细胞,对其生物学特性进行研究。间充质和上皮的相互作用是体外构建组织工程皮肤的基础,目前有关单一间充质细胞类型在组织工程皮肤制备中对表皮的形态发生研究较多,但是混合间充质细胞对表皮形态发生的作用未见报道,在获得人脂肪组织来源干细胞基础上,研究脂肪组织来源干细胞与真皮成纤维细胞的混合体对表皮形态发生的影响。方法:采用胶原酶消化人脂肪组织,低密度接种获得人脂肪组织来源干细胞,流式细胞仪检测细胞周期和细胞表面分子标志物,通过诱导分化检测人脂肪组织来源干细胞的多向分化潜能,在验证获取的人脂肪组织来源干细胞是间充质干细胞的基础上,进一步分离真皮成纤维细胞和角质形成细胞,MTT法观察脂肪组织来源干细胞与真皮成纤维细胞的混合体(细胞数量比为1:1)的条件培养液对角质形成细胞增殖能力的影响,并通过酶联免疫吸附试验(ELISA)对条件培养液成分进行检测,在此基础上以脂肪组织来源干细胞与真皮成纤维细胞的混合体与胶原凝胶复合构建组织工程真皮,并在其上接种角质形成细胞,组织学和形态测量学以及透射电子显微镜观察表皮形态发生的情况。结果:分离培养的人脂肪组织来源干细胞具有自我更新的能力,表达间充质干细胞的表面标志物,具有向脂肪、骨和软骨等多个谱系分化的能力;脂肪组织来源干细胞与真皮成纤维细胞的混合体的条件培养液可以促进角质形成细胞的增殖,ELISA检测其中富含肝细胞生长因子(HGF)和角质细胞生长因子(KGF也称为FGF7),在以脂肪组织来源干细胞与真皮成纤维细胞的混合体复合胶原为真皮构建的组织工程皮肤模型中,组织学分析表明,角质形成细胞在脂肪组织来源干细胞与真皮成纤维细胞的混合体上广泛增殖,并形成了较厚且分化良好的表皮层结构,并在表皮层的棘细胞层中检测到角蛋白10(表皮分化标记物)阳性,透射电镜(TEM)观察可见桥粒和半桥粒样结构,形态学测量所形成的表皮厚度与天然皮肤接近。结论:成功分离了人脂肪组织来源干细胞,其与真皮成纤维细胞混合有利于双层组织工程皮肤的构建,在较短的时间内构建出形态结构良好类似于天然皮肤的组织工程皮肤,提示脂肪组织来源干细胞在组织工程皮肤的构建中发挥着重要的作用。 3基于人脂肪组织来源干细胞和可降解明胶微球的工程化脂肪组织构建 目的:以EDC交联的明胶微球为支架,人脂肪组织来源干细胞为种子细胞,生物反应器进行三维培养并进行成脂诱导,构建工程化的脂肪组织。方法:制备明胶微球,并检测其细胞相容性和组织相容性,将PKH26标记的人脂肪组织来源干细胞接种于明胶微球上在旋转培养系统(RCCS)中培养,观察细胞在明胶微球上的生长状态,并对脂肪组织来源干细胞与明胶微球复合体进行成脂诱导7天,移植到免疫缺陷小鼠皮下,4周时取材检测细胞存活情况和脂肪形成。结果:经EDC交联的明胶微球均呈圆球体,大小比较均匀,粒径在75μm-150μm,脂肪组织来源干细胞在微球上生长状态良好,植入大鼠皮下,植入部位未见明显红肿,伤口无红肿、渗液等炎性反应,组织学观察植入明胶微球局部无明显炎症细胞浸润。旋转培养系统中,脂肪组织来源干细胞在明胶微球上吸附均匀,生长良好,经过体外成脂诱导7天后,在mRNA水平可检测到脂肪细胞相关基因PPAR-γ和C/EBPα,移植入免疫缺陷小鼠皮下,无论是对照组(未诱导)还是实验组(成脂诱导)在术后4周时都可以检测到PKH26标记的脂肪组织来源干细胞在微球周边存活,经成脂诱导7天的脂肪组织来源干细胞和明胶微球聚合物在裸鼠皮下可以形成脂肪样组织。结论:明胶是一种具有较好的细胞亲和性、生物相容性和生物可降解性的天然高分子材料,利用明胶制备明胶微球,以脂肪组织来源干细胞为种子细胞,在旋转培养系统中进行三维培养,可以为临床使用提供一种工程化脂肪组织填充物。
[Abstract]:Injuries, infections, diabetic ulcers, and tumor resection often cause soft tissue defects such as skin and fat. The problem of repairing soft tissue defects has long plagued clinicians. At present, there are various shortcomings in the clinical treatment methods, with the development and application of tissue engineering and stem cell technology and the use of stem cells. The construction of soft tissue, such as skin and fat, is the most promising way to repair soft tissue defects. The stem cells derived from adipose tissue are adult mesenchymal stem cells derived from adipose tissue, which have the ability of self renewal and multidifferentiation, and are easy to obtain, have large material, small damage, and rapid cell proliferation. The stem cells are the ideal seed cells for tissue engineering. First, we use the isolated SD rat adipose tissue derived stem cells to prove that it has the ability to promote wound healing in the skin defect model of rats, and further separate the stem cells from the human adipose tissue and make fiber from the dermis on this basis. The function of human adipose tissue derived stem cells in the preparation of tissue engineered skin was investigated by the mixed simulation of dermal microenvironment, and the adipose tissue derived stem cells were combined with the injectable gelatin microspheres to prepare the engineered adipose tissue. The tissue engineered skin and fat based on the stem cells derived from adipose tissue were explored to repair the tissue engineering skin and fat. A new strategy for soft tissue defects.
This subject is mainly studied in the following aspects:
Application of 1 rat adipose tissue derived stem cells in skin wound healing
Objective: to isolate and culture the stem cells from the adipose tissue of SD rats in vitro, to study the biological characteristics of the stem cells, and to construct tissue engineering dermis in collagen gel to repair the skin defect of rats, and to explore the feasibility of it as a tissue engineering skin seed cell. Methods: to isolate the groin adipose tissue of the SD rats and obtain the collagenase digestion. The stem cells were derived from the adipose tissue, and the growth characteristics of the cells were detected by MTT. Flow cytometry was used to detect the cell cycle and surface molecular markers. The multidirectional differentiation potential of stem cells derived from adipose tissue was detected by induction of differentiation. On the basis of verifying the derived stem cells of the adipose tissue derived from mesenchymal stem cells, Hoechst33 342 fluorescent markers were mixed with collagen gel to prepare tissue engineering dermis, which was used to repair the skin defect of rats. It was randomly divided into 3 groups: adipose tissue derived stem cell collagen transplantation group (ADSC+Col), simple collagen transplantation group (Col), blank control group (Cont.), and the repair effect through histology and healing speed. Adipose tissue derived stem cells have the ability to proliferate, express the surface markers of mesenchymal stem cells, and have the ability to differentiate into multiple lineages, such as fat, bone and nerve. In the compound collagen gel repair of skin defects, the adipose tissue derived stem cell collagen graft group (ADSC+Col), simple collagen transplantation group (Col), blank The wound healing time of group (Cont.) was (14.3 + 1.7), (16.9 + 2.5) and (21.2 + 4.2) days, with significant difference (P0.05). Histological observation showed that the adipose tissue derived stem cells were combined with collagen transplantation group, and a large number of fibroblasts and dermal collagen matrix were found to grow into the wound surface under light microscope, and the epithelium covered the wound, and the PNP like structure had been formed. But there was no skin appendage structure such as hair follicle and sweat gland, and simple collagen transplantation group, the epithelium crawled but not completely covered the wound, and there were more microvascular in the blank control group, but the speed of the epithelium crawled slowly, and the wound was not covered. The blue fluorescence was visible after 2 weeks after the transplantation of Hoechst33342 fat tissue stem cells combined with collagen transplantation. It was proved that the stem cells from the adipose tissue derived from the transplanted stem cells survived the wound. Conclusion: the stem cells derived from the adipose tissue derived from the adipose tissue were successfully isolated and the tissue engineered dermis constructed by the stem cells of the adipose tissue, which could repair the soft tissue defects of the skin, promote the healing of the wound, and prove that the stem cells derived from the adipose tissue were used as tissue engineering skin. The feasibility of building seed cells in skin.
The role of 2 human adipose tissue derived stem cells in tissue engineering skin preparation
Objective: to isolate and culture human adipose tissue derived stem cells in vitro, and to study their biological characteristics. The interaction of mesenchymal and epithelial cells is the basis for the construction of tissue engineered skin in vitro. At present, there are more studies on the morphogenesis of the epidermis in the preparation of tissue engineering skin, but mixed mesenchymal cells are mixed. The effect of epidermal morphogenesis was not reported. On the basis of obtaining human adipose tissue derived stem cells, the effects of the mixture of adipose tissue derived stem cells and dermal fibroblasts on the morphogenesis of epidermis were studied. Methods: using collagenase to digest human adipose tissue, low density inoculation to obtain human adipose tissue derived stem cells, flow finer Cell cycle and cell surface molecular markers were detected by cytosgraph, and the multidirectional differentiation potential of stem cells derived from human adipose tissue was detected by induction of differentiation. On the basis of verifying the derived stem cells derived from human adipose tissue derived from mesenchymal stem cells, the dermis fibroblasts and keratinocytes were further separated, and adipose tissue was observed by MTT method. The effect of the conditioned medium of source stem cells and dermal fibroblasts (the number of cells to 1:1) on the proliferation of keratinocytes and detection of the composition of the conditioned medium by enzyme linked immunosorbent assay (ELISA), and on this basis, the mixture and collagen of the stem cells and the dermal fibroblasts were derived from the adipose tissue. Gel composite construction of tissue engineering dermis, and inoculating keratinocytes on it, histology and morphometry and transmission electron microscopy to observe the occurrence of epidermal morphogenesis. Results: the isolated human adipose tissue derived stem cells have the ability to renew themselves, to reach the surface markers of mesenchymal stem cells, and to have the direction of lipid. The ability to differentiate multiple lineages, such as fat, bone, and cartilage; the conditioned medium of a mixture of adipose tissue derived stem cells and dermal fibroblasts can promote the proliferation of keratinocytes. ELISA detection is rich in hepatocyte growth factor (HGF) and keratinocyte growth factor (KGF, also known as FGF7), and in the stem cells derived from adipose tissue. The histologic analysis of the tissue engineering skin model constructed by the mixture of the dermal fibroblasts and the complex collagen of the dermal fibroblasts showed that the keratinocytes proliferate extensively on the mixture of adipose tissue derived stem cells and dermis fibroblasts, forming a thicker and well differentiated epidermis and in the spinous layer of the epidermis. Detection of keratin 10 (epidermal differentiation marker) positive, transmission electron microscopy (TEM) observation of the structure of grained and half bridge particles, morphological measurement of the thickness of the epidermis close to the natural skin. Conclusion: the successful separation of human adipose tissue derived stem cells, and its mixed with dermal fibroblast cells is conducive to the construction of double-layer tissue engineering skin. The construction of tissue engineered skin similar to natural skin is constructed in a short period of time, suggesting that adipose tissue derived stem cells play an important role in the construction of tissue engineered skin.
3 construction of engineered adipose tissue based on human adipose tissue derived stem cells and degradable gelatin microspheres
Objective: EDC crosslinked gelatin microspheres were used as scaffolds. Human adipose tissue derived stem cells were seeded cells. The bioreactor was used for three-dimensional culture and lipid induction, and engineered adipose tissue was constructed. Methods: gelatin microspheres were prepared, and their cellular compatibility and tissue compatibility were detected, and PKH26 labeled human adipose tissue derived stem cells were connected. The growth state of the cells on gelatin microspheres was observed on gelatin microspheres, and the growth state of the cells on gelatin microspheres was observed. The fat tissue derived stem cells and gelatin microspheres complex were induced for 7 days. The cells were transplanted subcutaneously into the immune deficient mice. The survival of the cells and the formation of fat were measured at 4 weeks. The results were as follows: gelatin crosslinked by EDC The microspheres were round spheroids, the size was more uniform, the size of the particles was 75 M-150 mu m. The stem cells of the adipose tissue derived from the microspheres were well grown on the microspheres. Adipose tissue derived stem cells were evenly adsorbed on gelatin microspheres and grew well. After 7 days of lipid induction in vitro, adipocyte related genes PPAR- gamma and C/EBP alpha could be detected at the mRNA level. The transplanted cells were transplanted subcutaneously into the immunodeficient mice. Both the control group (uninduced) or the experimental group (fat induced) could detect PKH at the end of the operation. 26 labeled adipose tissue derived stem cells survived the microspheres, and adipose tissue derived stem cells and gelatin microspheres could form adipose tissue subcutaneously in nude mice after 7 days of lipid induction. Conclusion gelatin is a natural polymer material with better cellular affinity, biocompatibility and biodegradability. Gelatin microspheres were prepared with the stem cells derived from adipose tissue as the seed cells, and three dimensional culture was carried out in the rotation culture system. It could provide a kind of Engineering fat tissue filling for clinical use.
【学位授予单位】：第四军医大学
【学位级别】：博士
【学位授予年份】：2011
【分类号】：R329
本文编号：2157659
[Abstract]:Injuries, infections, diabetic ulcers, and tumor resection often cause soft tissue defects such as skin and fat. The problem of repairing soft tissue defects has long plagued clinicians. At present, there are various shortcomings in the clinical treatment methods, with the development and application of tissue engineering and stem cell technology and the use of stem cells. The construction of soft tissue, such as skin and fat, is the most promising way to repair soft tissue defects. The stem cells derived from adipose tissue are adult mesenchymal stem cells derived from adipose tissue, which have the ability of self renewal and multidifferentiation, and are easy to obtain, have large material, small damage, and rapid cell proliferation. The stem cells are the ideal seed cells for tissue engineering. First, we use the isolated SD rat adipose tissue derived stem cells to prove that it has the ability to promote wound healing in the skin defect model of rats, and further separate the stem cells from the human adipose tissue and make fiber from the dermis on this basis. The function of human adipose tissue derived stem cells in the preparation of tissue engineered skin was investigated by the mixed simulation of dermal microenvironment, and the adipose tissue derived stem cells were combined with the injectable gelatin microspheres to prepare the engineered adipose tissue. The tissue engineered skin and fat based on the stem cells derived from adipose tissue were explored to repair the tissue engineering skin and fat. A new strategy for soft tissue defects.
This subject is mainly studied in the following aspects:
Application of 1 rat adipose tissue derived stem cells in skin wound healing
Objective: to isolate and culture the stem cells from the adipose tissue of SD rats in vitro, to study the biological characteristics of the stem cells, and to construct tissue engineering dermis in collagen gel to repair the skin defect of rats, and to explore the feasibility of it as a tissue engineering skin seed cell. Methods: to isolate the groin adipose tissue of the SD rats and obtain the collagenase digestion. The stem cells were derived from the adipose tissue, and the growth characteristics of the cells were detected by MTT. Flow cytometry was used to detect the cell cycle and surface molecular markers. The multidirectional differentiation potential of stem cells derived from adipose tissue was detected by induction of differentiation. On the basis of verifying the derived stem cells of the adipose tissue derived from mesenchymal stem cells, Hoechst33 342 fluorescent markers were mixed with collagen gel to prepare tissue engineering dermis, which was used to repair the skin defect of rats. It was randomly divided into 3 groups: adipose tissue derived stem cell collagen transplantation group (ADSC+Col), simple collagen transplantation group (Col), blank control group (Cont.), and the repair effect through histology and healing speed. Adipose tissue derived stem cells have the ability to proliferate, express the surface markers of mesenchymal stem cells, and have the ability to differentiate into multiple lineages, such as fat, bone and nerve. In the compound collagen gel repair of skin defects, the adipose tissue derived stem cell collagen graft group (ADSC+Col), simple collagen transplantation group (Col), blank The wound healing time of group (Cont.) was (14.3 + 1.7), (16.9 + 2.5) and (21.2 + 4.2) days, with significant difference (P0.05). Histological observation showed that the adipose tissue derived stem cells were combined with collagen transplantation group, and a large number of fibroblasts and dermal collagen matrix were found to grow into the wound surface under light microscope, and the epithelium covered the wound, and the PNP like structure had been formed. But there was no skin appendage structure such as hair follicle and sweat gland, and simple collagen transplantation group, the epithelium crawled but not completely covered the wound, and there were more microvascular in the blank control group, but the speed of the epithelium crawled slowly, and the wound was not covered. The blue fluorescence was visible after 2 weeks after the transplantation of Hoechst33342 fat tissue stem cells combined with collagen transplantation. It was proved that the stem cells from the adipose tissue derived from the transplanted stem cells survived the wound. Conclusion: the stem cells derived from the adipose tissue derived from the adipose tissue were successfully isolated and the tissue engineered dermis constructed by the stem cells of the adipose tissue, which could repair the soft tissue defects of the skin, promote the healing of the wound, and prove that the stem cells derived from the adipose tissue were used as tissue engineering skin. The feasibility of building seed cells in skin.
The role of 2 human adipose tissue derived stem cells in tissue engineering skin preparation
Objective: to isolate and culture human adipose tissue derived stem cells in vitro, and to study their biological characteristics. The interaction of mesenchymal and epithelial cells is the basis for the construction of tissue engineered skin in vitro. At present, there are more studies on the morphogenesis of the epidermis in the preparation of tissue engineering skin, but mixed mesenchymal cells are mixed. The effect of epidermal morphogenesis was not reported. On the basis of obtaining human adipose tissue derived stem cells, the effects of the mixture of adipose tissue derived stem cells and dermal fibroblasts on the morphogenesis of epidermis were studied. Methods: using collagenase to digest human adipose tissue, low density inoculation to obtain human adipose tissue derived stem cells, flow finer Cell cycle and cell surface molecular markers were detected by cytosgraph, and the multidirectional differentiation potential of stem cells derived from human adipose tissue was detected by induction of differentiation. On the basis of verifying the derived stem cells derived from human adipose tissue derived from mesenchymal stem cells, the dermis fibroblasts and keratinocytes were further separated, and adipose tissue was observed by MTT method. The effect of the conditioned medium of source stem cells and dermal fibroblasts (the number of cells to 1:1) on the proliferation of keratinocytes and detection of the composition of the conditioned medium by enzyme linked immunosorbent assay (ELISA), and on this basis, the mixture and collagen of the stem cells and the dermal fibroblasts were derived from the adipose tissue. Gel composite construction of tissue engineering dermis, and inoculating keratinocytes on it, histology and morphometry and transmission electron microscopy to observe the occurrence of epidermal morphogenesis. Results: the isolated human adipose tissue derived stem cells have the ability to renew themselves, to reach the surface markers of mesenchymal stem cells, and to have the direction of lipid. The ability to differentiate multiple lineages, such as fat, bone, and cartilage; the conditioned medium of a mixture of adipose tissue derived stem cells and dermal fibroblasts can promote the proliferation of keratinocytes. ELISA detection is rich in hepatocyte growth factor (HGF) and keratinocyte growth factor (KGF, also known as FGF7), and in the stem cells derived from adipose tissue. The histologic analysis of the tissue engineering skin model constructed by the mixture of the dermal fibroblasts and the complex collagen of the dermal fibroblasts showed that the keratinocytes proliferate extensively on the mixture of adipose tissue derived stem cells and dermis fibroblasts, forming a thicker and well differentiated epidermis and in the spinous layer of the epidermis. Detection of keratin 10 (epidermal differentiation marker) positive, transmission electron microscopy (TEM) observation of the structure of grained and half bridge particles, morphological measurement of the thickness of the epidermis close to the natural skin. Conclusion: the successful separation of human adipose tissue derived stem cells, and its mixed with dermal fibroblast cells is conducive to the construction of double-layer tissue engineering skin. The construction of tissue engineered skin similar to natural skin is constructed in a short period of time, suggesting that adipose tissue derived stem cells play an important role in the construction of tissue engineered skin.
3 construction of engineered adipose tissue based on human adipose tissue derived stem cells and degradable gelatin microspheres
Objective: EDC crosslinked gelatin microspheres were used as scaffolds. Human adipose tissue derived stem cells were seeded cells. The bioreactor was used for three-dimensional culture and lipid induction, and engineered adipose tissue was constructed. Methods: gelatin microspheres were prepared, and their cellular compatibility and tissue compatibility were detected, and PKH26 labeled human adipose tissue derived stem cells were connected. The growth state of the cells on gelatin microspheres was observed on gelatin microspheres, and the growth state of the cells on gelatin microspheres was observed. The fat tissue derived stem cells and gelatin microspheres complex were induced for 7 days. The cells were transplanted subcutaneously into the immune deficient mice. The survival of the cells and the formation of fat were measured at 4 weeks. The results were as follows: gelatin crosslinked by EDC The microspheres were round spheroids, the size was more uniform, the size of the particles was 75 M-150 mu m. The stem cells of the adipose tissue derived from the microspheres were well grown on the microspheres. Adipose tissue derived stem cells were evenly adsorbed on gelatin microspheres and grew well. After 7 days of lipid induction in vitro, adipocyte related genes PPAR- gamma and C/EBP alpha could be detected at the mRNA level. The transplanted cells were transplanted subcutaneously into the immunodeficient mice. Both the control group (uninduced) or the experimental group (fat induced) could detect PKH at the end of the operation. 26 labeled adipose tissue derived stem cells survived the microspheres, and adipose tissue derived stem cells and gelatin microspheres could form adipose tissue subcutaneously in nude mice after 7 days of lipid induction. Conclusion gelatin is a natural polymer material with better cellular affinity, biocompatibility and biodegradability. Gelatin microspheres were prepared with the stem cells derived from adipose tissue as the seed cells, and three dimensional culture was carried out in the rotation culture system. It could provide a kind of Engineering fat tissue filling for clinical use.
【学位授予单位】：第四军医大学
【学位级别】：博士
【学位授予年份】：2011
【分类号】：R329
【参考文献】
相关期刊论文 前1条
1 ;Full-thickness tissue engineered skin constructed with autogenic bone marrow mesenchymal stem cells[J];Science in China(Series C:Life Sciences);2007年04期
，本文编号：2157659
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