Transdifferentiation of adipose-derived stem cells into keratinocyte-like cells: engineering a stratified epidermis.
Transdifferentiation of adipose-derived stem cells into keratinocyte-like cells: engineering a stratified epidermis.
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DOI：
10.1371/journal.pone.0080587
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Galiano RD
中科院分区：
文献类型：
--
作者：
Chavez-Munoz C;Nguyen KT;Xu W;Hong SJ;Mustoe TA;Galiano RD
皮肤再生是组织工程领域中的重要研究领域，特别是对于涉及大面积皮肤损失的情况，其中当前的治疗不能诱导令人满意的永久替代。人脂肪源性干细胞（ASC）已被证明在体外分化成间充质谱系和非间充质谱系，证实了它们的转分化能力。这种多功能的分化潜力，加上其易于收获，使ASC处于基于干细胞的治疗的前沿。在这项研究中，我们假设ASC也有能力转分化成角质形成细胞样细胞，并能够进一步工程化的复层表皮。成功地从脂肪抽吸物中分离ASC并进行细胞分选（FACS）。分选后，将ASC与人角质形成细胞或角质形成细胞条件培养基共培养。经过14天的潜伏期，ASC形成了人类角质形成细胞特有的多边形鹅卵石形状。Western blot和q-PCR分析显示，在这些角质形成细胞样细胞（KLC）中存在特异性角质形成细胞标志物，包括细胞角蛋白-5、外皮蛋白、聚丝蛋白和分层蛋白; ASC中不存在这些标志物。为了进一步评估KLC是否能够类似于人角质形成细胞分层，将ASC接种在人脱细胞真皮的顶部，并在EGF和高Ca 2+浓度存在或不存在的情况下培养。组织学分析表明，在EGF和高钙的存在下培养时，在正常皮肤中观察到的分层结构相似。此外，免疫组织化学分析显示存在角质形成细胞标记物，如外皮蛋白，细胞角蛋白-5和10。总之，本研究首次证明了ASC具有转分化为KLC并设计成分层表皮的能力。这项研究表明，脂肪组织是潜在的角质形成细胞的一个容易获得的和可访问的来源，特别是对于严重的伤口，包括身体的大表面积和需要迅速上皮化。
Skin regeneration is an important area of research in the field of tissue-engineering, especially for cases involving loss of massive areas of skin, where current treatments are not capable of inducing permanent satisfying replacements. Human adipose-derived stem cells (ASC) have been shown to differentiate in-vitro into both mesenchymal lineages and non-mesenchymal lineages, confirming their transdifferentiation ability. This versatile differentiation potential, coupled with their ease of harvest, places ASC at the advancing front of stem cell-based therapies. In this study, we hypothesized that ASC also have the capacity to transdifferentiate into keratinocyte-like cells and furthermore are able to engineer a stratified epidermis. ASC were successfully isolated from lipoaspirates and cell sorted (FACS). After sorting, ASC were either co-cultured with human keratinocytes or with keratinocyte conditioned media. After a 14-day incubation period, ASC developed a polygonal cobblestone shape characteristic of human keratinocytes. Western blot and q-PCR analysis showed the presence of specific keratinocyte markers including cytokeratin-5, involucrin, filaggrin and stratifin in these keratinocyte-like cells (KLC); these markers were absent in ASC. To further evaluate if KLC were capable of stratification akin to human keratinocytes, ASC were seeded on top of human decellularized dermis and cultured in the presence or absence of EGF and high Ca2+ concentrations. Histological analysis demonstrated a stratified structure similar to that observed in normal skin when cultured in the presence of EGF and high Ca2+. Furthermore, immunohistochemical analysis revealed the presence of keratinocyte markers such as involucrin, cytokeratin-5 and cytokeratin-10. In conclusion this study demonstrates for the first time that ASC have the capacity to transdifferentiate into KLC and engineer a stratified epidermis. This study suggests that adipose tissue is potentially a readily available and accessible source of keratinocytes, particularly for severe wounds encompassing large surface areas of the body and requiring prompt epithelialization.
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DOI：
10.1097/00004630-199920060-00006
发表时间：
1999-11-01
期刊：
JOURNAL OF BURN CARE & REHABILITATION
影响因子：
--
作者：
Boyce, ST;Kagan, RJ;Warden, GD
通讯作者：
Warden, GD
影响因子：
4
作者：
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通讯作者：
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影响因子：
--
作者：
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通讯作者：
Hedrick, Marc H
影响因子：
4
作者：
Braun, Julian;Kurtz, Andreas;Dong, Jun
通讯作者：
Dong, Jun
影响因子：
3.7
作者：
Ecker, RC;Rogojanu, R;Steiner, GE
通讯作者：
Steiner, GE